December 2012

Transcription

December 2012

Volume 25 Number 12
BioPharm International
BioPharm
INTERNATIONAL
December 2012
The Science & Business of Biopharmaceuticals
DECEMBER 2012
www.biopharminternational.com
Employment Survey
I
Glycan Analysis
I
JOB SECURITY IN
A CHANGING
BIOPHARMA
ENVIRONMENT
Container-Closures
RESULTS FROM
OUR ANNUAL
EMPLOYMENT
SURVEY
Volume 25
PEER-REVIEWED:
Number 12
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BioPharm
I N T E R N AT I O N A L
The Science & Business of Biopharmaceuticals
EDITORIAL
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BioPharm
I N T E R N AT I O N A L
Contents
Volume 25 Number 12
December 2012
BioPharm International integrates
the science and business of
biopharmaceutical research, development,
and manufacturing. We provide practical,
peer-reviewed technical solutions
to enable biopharmaceutical professionals
to perform their jobs more effectively.
PEER-REVIEWED FEATURES
FEATURES
MEMBRANE SCALE-DOWN
A New Scale-Down Membrane
Adsorber Device for Process
Development and Validation
SURVEYS
Job Security in a Changing
Biopharma Environment
ON THE WEB
Social Media
Amy Ritter
Nathalie Frau, Martin Leuthold, Amit Mehta,
Kome (Kevin) Shomglin, and Rene Faber
Results from the 2012 employment survey. 12
The development of an ultra scale-down anion
exchange membrane adsorber.
18
Optimizing Global
Biopharmaceutical Operations
Through Risk Mitigation and
Management
Follow us on Twitter@BioPharmIntl
CONTAINER CLOSURES
Closures for Pharmaceutical
Preparations: A Review of
Design and Test Considerations
Tim Sandle
The author examines the use of closures
for products intended for injection.
32
Phil Kaminsky, Jiyang Liu, and Julia Olsen-Claire
A UC Berkeley survey provides insight
into biopharma’s risk concerns and strategies. 38
GLOBAL MARKETS
Navigating Emerging Markets
Jill E. Sackman
ANNIVERSARY
RETROSPECTIVE
A 25-Year Retrospective
on Computer Validation
An introduction to a new series on
manufacturing within global markets.
46
Join our BioPharmInternational Group
BioPharm Bulletin
Subscribe to the one industry
newsletter focused on the
development and manufacturing
of biotech drugs and vaccines.
Catch up on regulatory
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industry deals & more.
biopharminternational.com/subscribe
8
Cover: Atomic Imagery/Getty Images
COLUMNS AND DEPARTMENTS
6 From the Editor
Keeping tabs on crucial
medicines should be part of
emergency-preparedness plans.
Angie Drakulich
7 Global News
10 Regulatory Beat
White House and Congress
likely to struggle over funding.
Jill Wechsler
48 Boot Camp: Tech Guide
NIBRT’s Pauline Rudd on what
to expect when performing
glycan analysis.
52 Bioanalytical Best Practices
Preparation of biological
samples for chromatographic
analyses.
Roger N. Hayes
55 Final Word
Can postapproval FDA filings
immunize pharma companies
from patent lawsuits?
Kevin Murphy
and Andrew Nason
57 Ad Index/Marketplace
BioPharm InternationalJTTFMFDUJWFMZBCTUSBDUFEPSJOEFYFEJOrBiological Sciences Database (Cambridge Scientific Abstracts)rBiotechnology
and Bioengineering Database (Cambridge Scientific Abstracts)rBiotechnology Citation Index (ISI/Thomson Scientific)rChemical Abstracts (CAS)
rŞScience Citation Index Expanded (ISI/Thomson Scientific)rWeb of Science (ISI/Thomson Scientific)
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4
BioPharm International www.biopharminternational.com December 2012
Beat your best score
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Converting a promising API to a final drug can be as
tricky as hitting a hole-in-one, with poor solubility as
the biggest challenge. EMD Millipore helps you to
turn this challenge into a solution, e. g., with Meglumine,
our API grade counterion in ICH/Q7 quality. Along
with products for solid dispersions, we offer innovative
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formation to make your pipeline more efficient. Find
out how to beat your best score at
www.emdmillipore.com/bioavailability
EMD Millipore is a division of Merck KGaA, Darmstadt, Germany
From the Editor
When Disaster Strikes at Home
T
Angie Drakulich is the
editorial director of
BioPharm International.
Keeping tabs on
crucial medicines
should be part of
consumers’ and
manufacturers’
emergencypreparedness plans.
6
he editorial offices of BioPharm International are based in New Jersey, so when
Hurricane Sandy hit last month, we were all affected. Flooding and high winds
destroyed many coastline homes along with well-known destinations across
Atlantic City, the Jersey Shore, and the neighboring Manhattan boroughs. Suburban
communities and businesses (including many US biopharma manufacturing headquarters) across the state were without power for days and schools were shutdown for
a week or more in several counties. President Obama declared the state, along with
New York, a major disaster area. The entire ordeal was quite surreal, and our team
considers ourselves to be lucky that we made it through unscathed. Our thoughts go
out to those who are still recovering from the storm and trying to get back to normal.
Many common questions arose during the hurricane and its aftermath, including how long food products could be considered safe to eat without refrigeration and
where the closest open gas station was to refill generators. Pertaining to our industry
were questions about medications that required refrigeration (primarily injectables
and liquids), prescriptions that needed refilling (many doctors’ offices and drugstores
were closed due to flooding and power outages), and drug products that had gotten
wet or been lost in the storm.
Fortunately, FDA has a consumer webpage devoted to drug safety after a natural
disaster, whether it be exposure to fire, unsafe water (key for drugs that have to be
reconstituted), or lack of refrigeration (www.fda.gov/Drugs/EmergencyPreparedness/
ucm085200.htm). Certain life-saving drugs (e.g., insulin) can be used even if not cold
as long as they are not past their expiration date, says the agency webpage. There is
also FDA information on medical devices and their safety during a crisis, and even an
agency guidance aimed at sales representatives on how to deal with lost or stolen drug
samples in the aftermath of a disaster (looting in general was quite prevalent after
Hurricane Sandy).
There are also some rules of thumb to follow in any emergency-preparedness plan.
Many patient advocates recommend having on hand a 30-day supply of regularly
used medicines (both prescription and over the counter), as well as extra quantities of
devices needed to administer medications. But as a consumer, it seems there should
be even more information available for those unfortunate times when something out
of human control affects the medications we need. As an asthmatic, for example, I
was concerned about my inhaler’s pending expiration date and not being able to get a
refill without having to go the emergency room, which by the way, would have been
impossible given that the end of my street had a 60-foot tree and power lines across it.
Drug labels are already quite lengthy, but it may be worthwhile to add a few more
details. For instance, language may focus on how long a cold-chain medicine can go
without refrigeration, or how long a drug exposed to excessive heat can be considered
safe, or the reasons behind drug-product expiration dates. Some of these details may
be provided in medication guides that come with drug products, but even those of us
in the pharma industry know that the majority of end users do not read those packets
in full or keep them. This is probably an area where FDA and pharmacists can do a
better job educating consumers about the importance of medication guides.
Another piece of information worth including on labels may be where to find
information during emergency situations. As so many discovered during Hurricane
Sandy, we rely heavily on technology to get our information. Smart phones, in particular, became indisposable in the aftermath of the storm when power outages held
hostage traditional television, phone, and Internet access. Pharma companies could
consider, for example, having active Twitter feeds during emergencies so that people
can still access crucial information.
Various disasters and crises are bound to affect the global community in the
future. Now is the time to think about the crucial information we will need to have
on hand—and attached to our medications—when that next time strikes. z
Global News
Understanding Cellular Reprogramming
A publication in the Nov. 21, 2012
issue of Cell examines the mechanisms
underlying the reprogramming of somatic
to become induced pluripotent stem cells
(IPSCs). IPSCs have engendered much
excitement as potential tools for disease
modeling or for regenerative medicine,
but the methods used to produce them
are inefficient and time consuming,
limiting their commercial potential.
The method pioneered by Shinya
Yamanaka of Kyoto University, which
earned him a Nobel Prize, involves
adding four transcription factors, Oct4,
Sox2, Klf4, and c-Myc, to a somatic
cell, usually a skin cell (i.e., fibroblast).
A team of scientists from the University
of Pennsylvania looked at where on the
chromatin those factors were bound 48
hours after transfection to understand
the sequence of events that leads to
reprogramming. They found that Oct4,
Sox2, and Klf4 bound enhancer regions of
the chromatin distant from the genes they
regulate. The authors suggest that these
transcription factors act to open closed
chromatin structures, allowing transcription
machinery to access the DNA. C-Myc
appeared to act by enhancing the binding
of the other factors to the chromatin.
The researchers also found large
regions of the genome where the
transcription factors would not bind at
48 hours, but which were activated at
a later time. The DNA-binding proteins,
called histones, associated with the
refractory regions were found to be
chemically modified with a modification
called H3K9me3. Moreover, blocking
the enzyme that produced the H3K9me3
modification was found to accelerate the
reprogramming process.
By understanding cellular reprogramming
at the genetic level, scientists will be able to
better control the process.
—Amy Ritter
Source: A. Soufi, G. Donohue, and
K. Zaret, Cell online, DOI:0.1016/
j.cell.2012.09.045, Nov.15, 2012.
Report from
South Korea
Four domestic companies filed
a suit against the Ministry of
Health and Welfare of South
Korea claiming that the recent
price cuts made by the ministry
have affected their businesses.
On April 1, 2012, drug prices were
reduced by an average 17% and
affected the prices of 6506 drugs across the board. The first cut, announced
in 2011, decreases the price of off-patented drugs 30%, with the price of the
first generic drug set at 60% of the price of the off-patented drug. Originally,
prices of off-patented drugs decrease by 20%, while the first generic version is
set at 68% of the off-patented drug. The second cut reduces the price of drugs
due to illegal rebates. The combination of the two cuts means that certain
drugs will undergo double-price reduction and the final price rate can be up
to 53.55%.
Pharmaceutical companies are crying foul because these cuts would have a
direct impact on their business profits even though the agency claimed that
this move would ensure market sustainability and eradicate the problem of
illicit rebates. In fact, a Sinhan Investment & Securities source has indicated
that the majority of companies experienced a 20% fall in profits since the
policy took effect.
In response to their declining fortunes, some companies have opted
to increase prices of their over-the-counter (OTC) drugs while others have
boosted their R&D expenditure. Domestic companies such as Dong-A
Pharmaceutical and LG Life Sciences have committed 22% and 19%, of net
sales on R&D, respectively.
Cher Boon Piang, an analyst for Asia Pacific Pharmaceutical and Healthcare
of Business Monitor International (Asia), says, “Given the price cuts, companies
may withdraw drugs that are not profitable. Local companies may even
move away from generics. There is also a shift towards biosmiliars that opens
opportunity for companies.” In October 2011, Dong-A Pharmaceutical joined
hands with Tokyo-based Meiji Seika Pharma to build a biosimiliar plant
in Songdo. Recently, local companies Yuhan Corp and Teregen ETEX are
collaborating to commercialize the provision of individual genome services.
The recent US-Korea free-trade agreement (FTA) has also crippled domestic
players as it contains provisions protecting the intellectual property rights of
original drug developers. For instance, the Korean agency has to inform original
manufacturers if there are companies looking to produce generic versions.
Companies are denied market approval if an objection is posed by original drug
manufacturers and when the claimed patent exists. It is also mandatory for the
generic manufacturer to provide safety and efficacy information to ensure that
the generic version does not infringe on the original one.
Perhaps a gradual price cut would have helped in alleviating pressures
faced by industry players in South Korea. Cher says, “In general, companies
face[d] both financial and time issues when two price cuts were introduced
in 2011. If price reduction is gradual and made known to companies in a
timeframe that prepares them for such reduction, these companies can
draft and implement strategies to minimize the impact of price reduction.
Moreover, it is easier to have short-term solutions against gradual price cuts
compared to a larger one-off reduction.”
December 2012
7
Kevin Forest/Getty Images
Discovery Pipeline
Global News
Cher points out that price cuts cannot be
the only solution to contain rising government
expenditures and it is also unfair to shift
the burden onto companies. Instead, the
government should look into ways to cut
expenditure by subsidizing only the necessary
and/or increase premiums. These strategies
may create a negative impression, but they
are essential if a country is not doing well
economically. In addition, the burden of
healthcare cost should rest on individuals
instead, he adds.
Clearly, the objectives of the government
and the pharmaceutical industry players
differ greatly. On one hand, the government
seeks to lower healthcare costs. On the other
hand, industry players are looking for ways
to maximize profits. Asked if a balance can
be struck between both parties, Cher says,
“The level of compromise is dependent on
the attractiveness of the market. Typically,
an attractive market allows the government
to push through its policies as companies
are willing to forgo higher profit margins in
exchange for sustainable growth over a
time period.”
The South Korean market is characterized
by its aging population and an affluent
population. Growth potential is limited as
it has evolved to a developed market and
industry players expect it to have established
regulations. Therefore, it makes sense that
industry players are pre-alerted of any policies
in the government’s agenda. For example,
industry players were informed in 2010 of the
price disclosure policy to take effect in 2012.
In Japan, industry players understand that it is
the usual practice that price cuts occur once
every two years.
Despite its fragmented market and the pricecut setback, South Korea is ranked among the
world’s top 12 with $8 billion annual sales. Cher
adds, “In the long run, it has the necessary
ingredients to continue with its successful
pharmaceutical industry, strong support for
innovation, the willingness of the private sector
to explore these innovative technologies
and the demographic profile also supports
increased drug usage. The conflict between
the government and industry will definitely
arise again in the future, but we believe the two
parties will reach a compromise. “
—Jane Wan is a freelance writer
based in Singapore
8
A 25-Year Retrospective
on Computer System
Validation
Throughout BioPharm International’s 25th anniversary year, we have looked back
at articles published in the first volume of the journal. This month, Sharon Strause,
an industry consultant, provides a look back at “Computer System Validation Part I:
Testing and Verification of Applications Software” by Leonard J. Goren.
Computer system validation has changed in the past 25 years as technology
has become more complex. The majority of the documentation requirements
and software tests Leonard Goren listed in his article, “Computer System
Validation Part I: Testing and Verification of Applications Software,” are still
applicable today. One of the key missing procedures required to execute
computer system validation well is the supplier management process,
which should include the requirements for a vendor audit. A vendor audit
determines the vendor’s capability of producing a software application that
can be validated for a regulatory company’s use. A vendor audit verifies
how the code was developed, documented, and tested in all stages of
the development process. A vendor audit determines the quality system
procedures that a vendor has in place to ensure a well-developed software
application. The audit determines how code is managed (i.e., configuration
management), reviewed, internally documented, tested for the use and
misuse of its software, and changed. The vendor audit captures how data
produced by the software meet the requirements document, which starts
the process of a vendor audit. CGMP requirements are established in the
company’s requirements document for the software application including
any audit trail functionality; how the code is managed from an infrastructure
perspective (e.g., local, wide area, and web networks); and disaster recovery
process (i.e., backup process and offsite code management). A thorough
vendor audit showing good development practices will then allow a company
to qualify the software’s installation on their equipment, and validate it for
their use according to the computer system validation procedures in place
at the company. If practices at the vendor are not quality capable, then a
company can either chose another vendor or know in detail the additional
requirements that would need to be completed to fulfill the requirements of
computer system validation. This could mean thoroughly testing the software
for accuracy of the stated requirements of the software as it is received before
ever beginning the actual “validation for use” that is in place at the company.
The other crucial document that must be in place prior to assessing any
vendor is the requirements document. Goren addresses the functional
requirements in his 1988 article, but today, a requirements document includes
functional, technical, and regulatory requirements for a software application.
A completed requirements document helps to determine the types of
applications that should be assessed and also the types of testing that may
be required to ensure workability of the software. It’s an important starting
point in the computer system validation process. Having a computer system
validation process that starts with requirements and includes a good process
for supplier management and auditing can help to minimize the validation of
an application or make it more complex. z
—Our Retrospective series has included updates on separations technology,
mammalian cell culture, industry perceptions, orphan drugs, mAbs, GMP training,
cleanroom management, and more. For a complete list of Retrospectives and their
original 1988 articles, visit BioPharmInternational.com/Retrospectives.
December 2012
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Email [email protected]
©Patheon Inc. All rights reserved. Published 11/12 PATH0214R2
Regulatory Beat
Research Policies Pose New
Challenge for US Administration
White House and Congress likely to struggle
over funding for bio/pharmaceutical regulation.
10
Executives at bio/pharmaceutical companies are watching
closely at how tax and budget
proposals will affect corporate
tax rates and investment.
commit to these new programs in a period of
political uncertainty.
SPENDING CUTS AHEAD
The dark cloud looming over all these programs is the year-end “fiscal cliff,” with nearly
$500 billion in tax increases and spending cuts
scheduled to begin Jan. 1, 2013 unless Congress
acts. Executives at bio/pharmaceutical companies are watching closely at how tax and budget proposals will affect corporate tax rates and
investment, as well as the specific funding for
FDA, the National Institutes of Health (NIH),
and other activities important to biomedical
innovation and healthcare coverage.
All sides acknowledge the crucial need to
reduce both public and private outlays for US
healthcare, and drug prices and reimbursement
are a prime target, particularly related to outlays for federal government health programs
and Medicare Part D. House Democrats have
pressed for added rebates on drugs purchased by
Medicare drug plans for low-income “dual eligible” seniors, which could total more than $100
million over 10 years. There also are budget proposals on the table to reduce federal spending
on drugs for federal government employees, as
well as other government health programs.
Digital Vision/Getty Images
T
he calm after the heated election battle
this year has been brief due to pressure on policymakers to tackle overwhelming budget issues. With Republicans
maintaining tight control over the House
of Representat ives but losing g rou nd in
the Senate, much depends on the abilit y
of President Obama to engineer some kind
of “fix” to the mounting deficit during the
year-end “lame duck” Congressional session.
Healthcare policy was a key point of dispute
during the election campaign, marked by
promises of better coverage and predictions of
soaring costs by both candidates. Now, scheduled funding cuts and significant tax increases
are expected to play a large role in shaping the
reform program.
The Obama v ictor y ended prospects of
wholesale repeal of the Affordable Care Act
(ACA). House Republicans will continue to
challenge various requirements of the healthcare legislation, but key provisions for pharmaceutical companies, such as rebates on drugs for
seniors in the Part D coverage gap and authorization for biosimilars, are unlikely to change.
More broadly, the promised expansion of coverage to some 30 million
prev iously uninsured A mer icans
will move forward, although with
consumers paying higher premiums
and cost-sharing to cover ever-rising healthcare costs. That sets the
stage for significant growth in the
market for brand-name drugs. The
Department of Health and Human
Jill Wechsler is BioPharm Services (HHS) is working hard to
International’s Washington editor, meet a host of deadlines and timeChevy Chase, MD, 301.656.4634, frames for establishing exchanges,
[email protected]. defining benefits, and expanding
Read Jill’s blogs at Medicaid, much of that involving
PharmTech.com/wechsler. states that have been reluctant to
Regulatory Beat
The Obama victory offers some
stability for FDA, as the agency
continues to implement the FDA
S a fet y & I n novat ion Ac t a nd
struggles to find a middle ground
between speeding untried new
medicines to patients and protecting the public from undue risk
and harm. Although there won’t
be a wholesale change in executive branch leadership, many top
administration officials are likely
to move on to other roles, and
extensive cuts in the 2013 budget could undermine many FDA
projects. An 8.2% cut in the FDA
budget, as proposed under the
sequester process, would reduce
FDA’s 2013 budget by $320 million and prompt the agency to lay
off approximately 1000 employe e s, accord i n g to con s u lt a nt
Steven Grossman, publisher of
FDA Matters. Even without such
a severe, across-the-board cut,
which could jeopardize FDA’s ability to collect user fees from pharmaceutical and medical device
ma ke r s, t he F DA budget w i l l
remain vulnerable to pressures to
reduce federal spending for some
years to come.
Severe reductions in NIH funding, moreover, would jeopardize
the pace of new drug and biotech
discovery and support for clinical research that is key to spurring innovation needed to fill the
depleted new drug pipeline. The
biomedical research community
is highlighting the importance of
both FDA and NIH in protecting
public health for patients at home
and around the world.
CONGRESS TACKLES
COMPOUNDING
Meanwhile, mounting deaths from
contaminated steroid injectables
made by a Massachusetts compounding pharmacy are focusing attention on the need for
broader FDA legal authority in
this and other areas. The need
to reauthorize animal-drug user
fees in 2013 is expected to provide a vehicle for legislation that
would better secure the prescription drug supply chain and also
address drug compounding oversight. (For more information on
this subject, view “Compounding
and FDA Regulation” on w w w.
BioPharmInternational.com.)
The ongoing fungal meningitis
outbreak had sickened more than
425 individuals and caused over 30
deaths, as of early November. Rep.
Edward Markey (D-MA) has proposed legislation to enhance FDA
oversight of compounding pharmacies, and lead House and Senate
committees held hearings right
after election day to address the
response by FDA and state regulators and to analyze actions by the
offending compounder, the New
England Compounding Center
(NECC). Markey’s bill clarifies
FDA’s right to inspect and regulate large compounders that qualify as drug manufacturers. Small
compounding pharmacies would
continue to operate under state
licensing, and FDA could issue
waivers to operators responding to
drug shortages and public health
crises. Compounded drugs have to
be labeled that they have not been
tested for FDA safety and efficacy
standards, and FDA has to publish
a list of unsafe or ineffective drugs
not suitable for compounding.
FDA regulation of compounders has been a thorny issue for
decades, as previous efforts by the
agency to impose stricter rules on
compounders have been struck
down by the courts. But the recent
crisis has reopened the debate
over the adequacy of state versus
federal regulation of pharmacies
and when compounding qualifies
as drug manufacturing.
Efforts by FDA and Massachuset ts reg u lators to shut dow n
N ECC and its sister company,
Ameridose, highlight the links
between drug shortages and compound ing. FDA Commissioner
Margaret Hamburg noted that
the agency is working hard to
minimize shortages in important
Ameridose products used in surgery and to prevent congestive
heart failure. Yet, former FDA
official Scott Gottlieb also commented that too-tight FDA regulations have led to shortages of
low-cost injectable drugs, prompting hospitals and patients to seek
a lte r nat ives f rom comp ou nders. Too-low reimbursement for
generic injectables also may limit
pharmaceutical industry interest
in producing these therapies, leading to shortages and greater reliance on compounders.
K V Pharma weighed in that
the NECC case illustrates FDA’s
error in permitting compounders
to continue to produce hydroxyprogesterone to prevent premature births after approving KV’s
Makena. State health agencies and
insurers have been opting for the
less costly compounded version,
but now may shift to the KV product to avoid exposure to possibly
unsafe compounded medicines.
There will be f urther debate
ove r how muc h a d d e d le g a l
authority FDA needs to deal with
compounders. Some agency critics complained that FDA did not
make full use of its existing legal
authority to regulate NECC follow i ng i n it i a l u n s at i sf ac tor y
inspections. Republicans generally
oppose giving the agency stronger
legal powers, and compounding
pharmacies claim they are sufficiently regulated by state licensing boards. Pharmacists object
to the Markey bill for imposing
too-broad FDA regulation of compounding that could block patient
access to needed medicines and
further overtax FDA. It’s a costversus-safety issue, and a challenge to find a compromise that
passes muster. ◆
December 2012 www.biopharminternational.com BioPharm International
11
Employment Survey
Job Security in a Changing
Biopharma Environment
Amy Ritter
Results from
the 2012
employment
survey.
R
eader responses to BioPharm
International’s annual employment survey suggest that the
mergers and acquisitions that
dominated the news a few years ago are
slowing. However, bio/pharmaceutical
companies still face pressure to run leaner
businesses and to see a better return on
investment from their R&D divisions.
Development of large-molecule therapeutics is an area of intense interest for large
pharma and small biotech companies, but
biologics are expensive to develop, which
affects companies’ bottom lines. In addition, bio/pharmaceutical companies face
pricing pressure from cost-conscious payers and from developing countries determined to hold the line on drug prices.
The industry continues to adapt to this
challenging business environment, and
12
these challenges cannot help but affect
pharma employees.
When readers were asked how secure
they felt in their positions, fewer than last
year said they felt less secure—33%, compared with 38% in 2011. While this seems
encouraging, readers did not say they felt
more secure. Instead, the largest group
of respondents (49%) said they felt about
the same as last year. It seems, then, that
pharma employees are becoming accustomed to the new, more fluid business
environment. Is insecurity becoming the
new normal? Perhaps, but most readers felt
confident they would be able to find a new
job if they had to, and readers continue
to derive satisfaction from the intellectual
stimulation and challenging projects associated with their jobs. The following pages
highlight key results from the survey.
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DynaPro® Plate Reader II. Automated dynamic
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96 or 384 or 1536 well plates, and now with an
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Employment Survey
Has your job become more global
in nature (i.e., more offices/sites
and/or partners in other countries)
over the past two years?
How secure do you feel in your job compared
with last year?
I feel more secure now
I feel less secure now
No change
2011
23.3%
2012
48.5%
38.6%
30.6%
18.6%
58.5%
32.9%
38.1%
10.9%
More global interactions
If it was necessary for you to change jobs this year,
how would you assess the job market?
15.6%
45.1%
Fewer global interactions
About the same
It would be straightforward to find a job
Has having offices/partners in
other countries made your job:
comparable to the one I have now.
It would take a while, but I would be able to find a job comparable
38.9%
to the one I have now.
15.9%
It would be straightforward to find a job, but it probably wouldn’t
23.4%
I would have to search hard, and be prepared to take what I could
53.4%
be as good as the one I have now.
get.
Within the past year,
has your workload
increased, decreased,
or stayed the same?
7.8%
2011
2012
More difficult
Less difficult
No different than working with local partners
71.1%
57.1%
How do you communicate with
other offices or partners in other
countries (check all that apply)?
Increased
Decreased
Stayed the same
98.4%
Teleconference 81.9%
In-person visits 56%
Email
35.3%
24.1%
7.6%
4.8%
Skype or other streaming video
24.9%
Employment Survey
This is the main reason
I come to work
In the past two years, have you
been through a merger, acquisition,
downsizing or restructuring?
39.8%
Intellectual
stimulation
Challenging
projects
38.9%
YES NO
2012
2011
$
MEDIAN SALARIE
US (USD)
Canada (CAD)
44.6% 55.4%
57.7% 42.3%
Salary
Professional
advancement
110,000
72,500 (72,535 USD)
What is your prediction for your company’s
business prospects in the coming year?
Business will improve.
Business will decline.
No significant change expected.
28.6% 32.1%
30.3%
50.2%
16.3%
33.6%
I would change jobs
for this alone.
In your view, what is the general outlook for the bio/pharmaceutical
industry in the short- and long-term?
51.8%
Business will improve.
11.2%
14.9%
*Due to rounding, some
percentages may not
add up to 100%. Some
questions allowed
multiple answers.
3.6%
2.6%
0.3%
15.5%
Business will decline.
Business will improve overseas, but not domestically.
Business will improve domestically, but not overseas.
Business will decline domestically, but not overseas.
Business will decline overseas, but not domestically.
No significant change expected.
Job
security
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Nathalie Frau, Martin Leuthold, Amit Mehta, Kome (Kevin) Shomglin, and Rene Faber
ABSTRACT
Anion exchange membrane chromatography (AEX) is an attractive alternative to flow-through anion exchange
column chromatography. Replacing AEX column chromatography with AEX membrane chromatography provides
similar output but at a much higher load density, usually greater than 10 kg/L of membrane. The commercially
available scale-down model, Sartobind nano, which has a 1 mL membrane volume, requires a significant amount of
material for process development and validation whereas a relatively small amount of material is typically available
during early clinical development. To overcome this limitation, an ultra scale-down device, Sartobind pico, was
developed to reduce material consumption and validation cost. In this article, the development of the new ultra
scale-down device is detailed and scalability to Sartobind nano and to a large-scale capsule are demonstrated.
Studies using model proteins and industrially relevant monoclonal antibody feedstock are described. The new ultra
scale-down device, Sartobind pico, enables process development, characterization, and validation with scalability to
large-scale membrane chromatography devices while reducing sample consumption, time, and cost.
Nathalie Frau, PhD*, is a senior scientist
in R&D process technologies at Sartorius
Stedim North America, Bohemia NY; Martin
Leuthold, PhD, is a scientist in R&D product
development at Sartorius Stedim Biotech,
Goettingen, Germany; Amit Mehta, PhD, is a
senior engineer in purification development
and Kome (Kevin) Shomglin, PhD, is a senior
research associate in purfication development
at Genentech, South San Francisco, CA; and
Rene Faber, PhD, is vice-president, R&D
process technologies at Sartorius Stedim,
North America, Bohemia NY. *To whom
correspondence should be addressed,
[email protected].
PEER-REVIEWED
Article submitted: Jul. 23, 2012.
Article accepted: Aug. 3, 2012.
18
A
nion-exchange (AEX) membrane chromatog raphy is
an attractive technology for
monoclonal antibody (mAb)
purification because of advantages
such as elimination of column packing and unpacking, higher throughput, smaller plant foot pr int, and
considerably less buffer consumption.
Compared with AEX resins, which
are typically loaded to approximately
100 g/L, AEX membranes can provide orders of magnitude higher loading capacity in flow-through mode
w it h adequate impu r it y remova l.
For example, Zhou et al. reported
greater than 3000 g/m 2 or 10.9 kg/L
load capacity with > 5 log reduction
value (LRV) for four different model
viruses (1). In another study, Zhou
et al. showed that a similar LRV for
X-MuLV could be obtained at a load
capacity of 13 kg/L and at flow rate of
600 cm/hr (2). Glynn et al. recently
described the evolution of Pfizer’s
antibody purification process from
three columns to two by replacing
the resin-based AEX chromatography step with a membrane adsorber
and increasing the load capacity of
this step by a factor of 100 (3). The
removal of process-related impurities
with AEX membrane adsorbers at high
load capacity and high flow rate has
also been published by Arunakumari
et al. (4). Lately, the authors demonstrated virus removal by membrane
adsorbers with a LRV greater than 4.5
and 4.4 for X-MuLV and MMV, respectively, at 20 kg/L mAb load capacity
(5). Mehta et al. showed that purity
and product quality comparable to
traditional three-column affinity processes can be achieved with a novel
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Erwin-Rentschler-Straße 21
88471 Laupheim / Germany
0HONEs%-AILINFO RENTSCHLERDE
www.rentschler.de
ALL FIGURES ARE COURTESY OF THE AUTHORS
Membrane Scale-Down
process using a nonaffinity capture step and
membrane-based technologies such as AEX
membrane adsorbers and high performance
tangential flow filtration (6).
It is thus well documented in the literature that an AEX membrane adsorber is a
powerful alternative to column chromatography and can facilitate development of
new purification strategies for downstream
processing in the biopharmaceutical industry (7). However, the high load capacity
achieved with membrane adsorbers in the
flow-through mode implies the need for a
significant amount of material for process
development with laboratory-scale devices.
For example, a load capacity of 10 kg/L
means that 10 g of material is required for
each experiment with a 1 mL laboratoryscale device. High material consumption
can be a limiting factor, particularly during
early stages of drug development where relatively small amount of material is typically
available. Reducing the virus validation cost
by minimizing the amount of virus spike
required is also of significant interest.
To overcome these limitations, a new
u lt r a s c a le - dow n me mbr a ne ad s or b e r
device, Sartobind pico (Sartorius Stedim
Biotech GmbH, G öt t i ngen, G er ma ny),
with a membrane volume of 0.08 mL has
been developed. The 12.5-fold lower membrane volume than the current laboratory-scale device, 1 mL Sartobind Nano,
significantly minimizes feedstock and virus
spike requirements for development, characterization, and validation studies. The performance of this device was evaluated using
model molecules and industrially relevant
mAb feedstock and was compared with the
current scale-down device, Sartobind nano.
Data demonstrating the scalability of the
new ultra scale-down device to a manufacturing-scale device are also presented.
MATERIALS AND METHODS
Devices
Sartobind pico, the new scale-down device
was provided by Sartorius Stedim Biotech
GmbH, Göttingen, Germany. The device
consists of 15 membrane layers with polypropylene sealing rings every 3 layers, and
is assembled into a molded polypropylene housing with luer lock connectors to
enable easy connection to a liquid chroma-
20
Table I: Key attributes of Sartobind pico and Sartobind nano.
Sartobind nano
Sartobind pico
Bed height (mm)
4
4
Membrane volume (mL)
1
0.08
Housing materials
Polypropylene
Polypropylene
Connectors
Luer-Lock
Luer-Lock
Flow path
Radial
Axial
Figure 1: A. Sartobind pico 0.08 mL. B. Sartobind nano 1 mL.
Figure 2: Sartobind pico device design.
Luer lock
Molded polypropylene housing
Stacked membrane design (5 x 3 layers)
tography system (see Figures 1 and 2). The
bed height of 4 mm is similar across the
entire Sartobind SingleSep family and the
frontal surface area of 20 mm² gives pico a
membrane volume of 0.08 mL. Sartobind
nano, (Sartorius Stedim Biotech GmbH,
Göttingen, Germany) with 15 layers, 36.4
cm 2 total surface area, and 1 mL membrane
volume was used as a reference device (see
Figure 1). The Sartobind nano has a radial
flow and is constructed in the same way
Setting New Standards:
Platform Solutions for High Quality Analytics
of Process Contaminants and Impurities
0N-DEMAND WEBCAST
Register free at www.biopharminternational.com/platform
EVENT OVERVIEW:
Monitoring biopharmaceutical product quality requires uncompromised analytics where high quality data combined with a fast time to
result enables efficient development and manufacturing processes.
This webinar will describe an integrated platform of rapid, highly
sensitive in-process testing methods that deliver industry-leading
solutions for contaminant and impurity detection and quantitation.
The methods include easy-to-use molecular technologies, including
quantitative PCR, DNA sequencing, and automated sample preparation that can be applied from cell line development to formulation.
Specific topics to be covered are:
In-process monitoring during cell-culture manufacturing
Using an integrated sample-to-results detection system for detection
of Mycoplasma, MMV and Vesivirus
High-throughput residual host cell DNA quantitation
An immunoassay platform for analysis of residual proteins
Key Learning Objectives:
Understand how and where rapid
molecular methods for highly sensitive
contaminant and impurity testing can be
applied
Learn strategies for using in-process
monitoring during cell-culture
manufacturing as an early warning signal
of a contamination event
Gain insight into an integrated analytical
testing platform that can help increase
throughput throughout the workflow
Who Should Attend:
This program is intended for scientists
working on or interested in learning more
about analytical methods for pharmaceutical contaminant and impurity testing in
such areas as:
Speaker:
Analytical Testing
Wesley Straub
Process Development
Senior Technical Product Specialist, Pharmaceutical
Analytics
Process Engineering
Life Technologies
Automation Engineering
Quality Control
Moderator:
Angie Drakulich
Managing Editor
Presented by
Manufacturing Science
Technical Services/Technology
Development
Sponsored by
For questions, contact Sara Barschdorf at [email protected]
Membrane Scale-Down
Figure 3: Normalized flow (MV/min) for Sartobind pico and nano devices
as a function of inlet pressure. MV is membrane volume.
80
70
Flow [MV/min]
60
50
40
30
20
10
0
0
0.5
1
1.5
2
2.5
3
3.5
Pressure [bar]
Pico 1
Pico 2
Nano 1
Nano 2
as process scale SingleSep capsules, which
assures direct scalability to manufacturing
scale capsules (7–11). The key attributes of
Sartobind pico and Sartobind nano are summarized in Table I .
The Sartobind SingleSep 10” capsule with
a membrane volume of 180 mL was used
to further confirm scalability. The devices
were assembled with a salt tolerant AEX
membrane, Sartobind STIC PA, consisting of
a polyallylamine ligand covalently coupled
to the cellulose membrane matrix (12).
Equipment
A l l l a b o r at o r y- s c a l e c h r o m at o g r ap hy
experiments with mAb feedstock, model
proteins, and model DNA were performed
using an ÄKTA Explorer FPLC system (GE
Healthcare Bio-Sciences Corp., Piscataway,
NJ, USA). The devices were connected to
the ÄKTA Explorer with standard tubing
and luer-lock connectors. A flow rate of
10 membrane volume (MV)/min was used.
Binding of endotoxin and bacteriophage
molecules was performed using a separate
experimental setup consisting of a peristaltic pump (Watson Marlow 302S), which
allowed proper cleaning of the system. To
determine flow rates, membrane adsorber
devices were connected to a pressure vessel filled with buffer or protein solution.
The filtrate volume was monitored using
a balance and the flow rates for different
pressures were calculated up to an inlet
pressure of 3 bar.
22
Model systems
Bovine serum albumin (BSA, Lot 50121326)
was purchased f rom K raeber GmbH &
Co. and salmon sperm DNA (DNA, Lot
8087) from Biomol. The protein throughput wa s deter m i ned usi ng γ -g lobu l i n
(Sigma, γ−globulin from bovine blood, Lot
STB0227K9). Endotoxin from Escherishia
coli (Lonza LPS E. coli 055:B5 N185 Lot
0 0 0 010 0778) w a s u s e d a s s t a n d a r d .
Bacteriophage ΦX174 (ATCC 13706-B1) was
produced in a 50 L disposable bioreactor
using the E. coli (ATCC 13706) expression
system. Subsequently, phage was purified,
concentrated, and sterile filtered by several
steps including a depth filtration cascade,
crossflow filtration, precipitation with polyethylene glycol, and centrifugation.
MAb feedstock
The mAb feedstock was obtained from pilotscale batches produced at Genentech (a member of the Roche Group). It was expressed
in mammalian cells and clarified to remove
insoluble impurities. The mAb was processed
through a protein A chromatography step
and further purified using a cation-exchange
chromatography step. Protein concentration
was approximately 11 g/L.
METHODS
Dynamic binding capacity
Each device was sanitized with 1 N NaOH
for 30 min at 10 M V/min followed by
equilibration with 150 MV binding buffer composed of 150 mM NaCl in 20 mM
Tris/HCl pH 7.3 ± 0.1, conductivity 16 mS/
cm. 150 MV of 1 g/L BSA in binding buffer or 0.1 g/L DNA in binding buffer were
loaded. All solutions used were prefiltered
with a 0.2 μm membrane filter. All steps
were performed at flow rate of 10 MV/min.
Breakthrough c ur ves were recorded by
measuring the extinction at 280 nm (protein) and 260 nm (DNA) using the ÄKTA
Explorer. To compare different devices the
void volume of the experimental setup
was determined by injection of acetone (2
%). The dynamic binding capacity at 10%
breakthrough was calculated as shown in
Equation 1,
DBC =
(V10% – Vv)* Ci
VM
[Eq. 1]
Membrane Scale-Down
Protein throughput
Each membrane adsorber device was sanitized with 1 N NaOH for 30 min at 10 MV/
min followed by equilibration with 100 MV
binding buffer composed of 150 mM NaCl
in 20 mM Tris/HCl pH 7.3 ± 0.1, conductivity 16 mS/cm. Protein throughput was determined using the pressure vessel filled with
a solution of 20 g/L γ-globulin in binding
buffer was used to determine the protein
throughput with the membrane adsorber
devices. The filtrate volume up to 1000 MV
was monitored at a constant pressure of 3
bar using a balance.
Chinese hamster ovary proteins clearance
Chinese hamster ovary proteins (CHOP)
clearance was determined using industrially
relevant mAb feedstock. Before loading the
MAb feedstock onto the membrane adsorber,
the membrane was equilibrated with 10 MV
of 50 mM Tris buffer at the appropriate pH.
The conductivity of this buffer was adjusted
by altering the concentration of sodium acetate. After equilibration, the mAb feedstock
was loaded onto the devices to a targeted load
density of 10 kg mAb/L of membrane at a
flow rate of 10 MV/min. Pool fractions were
collected during the experiment and analyzed for CHOP concentration.
Determination of log
reduction value of bacteriophages
Equipment and membrane devices were sanitized with 1 M sodium hydroxide for 30
minutes. Membrane devices were further
equilibrated with 300 MV of binding buffer. The ΦX174 phage solution with a titer
of 1.5x107 PFU/mL was prepared and loaded
onto the devices at a flow rate of 10 MV/
min. Flow-through fractions were collected
after 100 and 150 MV of load for quantitative analysis.
Endotoxin removal
Pump, tubing, and devices were treated
with 1 M sodium hydroxide for 30 minutes
at room temperature and at a flow rate of
10 MV/min before performing the experiment. Compatible vessels and materials
Figure 4: Filtrate flow rate at 3 bar constant initial pressure during loading
of 20 g/L γ-globulin protein solution. MV is membrane volume.
35
Flow [MV/(min*bar)]
where V10% is volume loaded at 10% breakthrough, Vv is void volume, Vm is membrane
volume, and ci is initial concentration.
30
25
20
15
10
5
0
0
5
10
Load density [kg/L]
Pico 1
Pico 2
Nano 1
15
20
Nano 2
were heated at 200 ∘C for 4 hours to destroy
naturally occurring endotoxins. After sufficient rinsing with reverse osmosis water,
the equilibration was performed with 300
MV of binding buffer. 150 MV of endotoxin
in binding buffer were loaded to the membrane at a flow rate of 10 MV/min. The flowthrough was divided into fractions of 50
MV each and was analyzed to determine the
endotoxin level.
ASSAYS
CHOP quantification
An ELISA was used for CHOP quantification. Samples containing CHOP were incubated in the wells, followed by incubation
w ith anti- CHOP antibodies conjugated
with horseradish peroxidase (HRP). The
HRP enzymatic activity was detected with
o-phenylenediamine, and the CHOP was
quantified by reading absorbance at 490
nm in a microtiter plate reader. Based on
the principles of sandwich ELISA, the concentration of peroxidase corresponded to
the CHOP concentration. The assay range
for the ELISA was typically 10–320 ng/mL,
with intra-assay variability of approximately
10%. CHOP values were reported in units
of ng/mL. CHOP values could be divided
by the mAb concentration and the results
reported in units of PPM (parts per million;
ng of CHOP/mg of mAb).
23
Membrane Scale-Down
Figure 5: Bovine serum albumin breakthrough curves for pico, nano, and
10” devices. MV is membrane volume.
The LRV was calculated using Equation 3,
LRV = log10 C0
CFT
1
[Eq. 3]
where c0 was the titer of the initial solution
and cFT the titer in the flow-through fraction.
0.8
C/C0
0.6
0.4
0.2
0
0
20
40
60
80
100
Load Volume [MV]
Pico 1
Pico 2
Nano 1
Nano 2
10” capsule 1
10” capsule 2
Bacteriophage ΦX174 quantification
Host organism E. coli was used for the detection of infectious ΦX174 phage particles.
E. coli cells were incubated on agar plates
(Soybean- Casein Digest Agar Medium–
Trypticase Soy Broth 211043), which served
as a base layer with nutrients. E. coli cells
multiplied rapidly and formed a bacterial
lawn. Phage particles infect the cells, causing
the lysis of E. coli host cells and producing
single circular, nonturbid areas called plaques
in the bacterial lawn. Each plaque represents
the lysis of a phage-infected bacterial culture
and is designated as a plaque-forming unit
(pfu), and used to quantitate the number
of infective phage particles in the culture.
Plaques must be clearly defined and samples
were then diluted several times (1:10) depending on the phage concentration. During the
study, 150 μL of the host cell solution (optical density 2–6) was mixed with 150 μL of
sample and top agar (1.3% Tryptikase Soy
Agar BD 211043) and the mixture was then
distributed to agar plates (4% Tryptikase Soy
Agar BD 211043 in 90 mm petri dishes) and
incubated for 18 to 24 hours at 37 ∘C. Plaque
forming units were counted and the titer of
the sample in PFU/mL (plaque forming units
per mL) was calculated using Equation 2,
Titer =
P
E
D
VSample
[Eq. 2]
where P is the number of plaques of all
countable dilutions, E is the sum of emphasis, D is the lowest evaluated dilution, and
VSample is the sample volume.
24
Endotoxin quantification
The endotoxin level was measured by the
kinetic chromogenic method test according to t he manufac t urer’s inst r uc t ions
(Limulus Amebocyte Lysate Chromogen,
Cha rles R iver endosa fe E ndoch rome -K
R1710K, Lot A4992L 10/2012). The quantification principle is based on coloration
caused by the contact of a sample containing endotoxin with a mixture of lysate
and chromogenic substrate. A β -glucan
blo c ke r w a s a d d e d ( L o n z a N19 0 L o t
0 0 0 0132199 01/11). D ur ing t he 1-hour
incubation the extinction coefficient was
measured continuously at 405 nm using a
temperature controlled (37 ∘C) plate reader
( Tecan Saf ire). T he react ion rate var ies
with endotoxin level and the samples were
quantified for endotoxin by comparing
t he results w it h t he calibrat ion ser ies.
The detection limit of the assay was 0.012
EU/mL. LRV was calculated similarly to
phage quantification by measuring the
endotoxin level of the initial solution El0
and t he level of endotox in in t he collected flowthrough fractions (ElFT) using
Equation 4 .
LRV = log10 EI0
EIFT
[Eq. 4]
RESULTS
Flow rate and protein throughput
Device geometry must allow for linear scalability through the entire device size range.
Pressure flow curves were generated with
the axial flow Sartobind pico and radial
f low Sartobind nano devices with data
shown in Figure 3. The normalized flow rate
(membrane volume (MV)/minute) increased
linearly with the increasing inlet pressure
and the flow rates were comparable, suggesting effective flow distribution and efficient
utilization of membrane area with both pico
and nano devices.
For a typical polishing application with
an AEX membrane adsorber, the load capac-
Membrane Scale-Down
Table II: Dynamic binding capacity (DBC) at 10% breakthrough using bovine serum albumin (BSA) and DNA model molecules.
BSA is bovine serum albumin.
Membrane
volume (mL)
10% DBC
BSA (g/L)
10 % DBC
DNA (g/L)
Pico 1
0.08
55.83
9.06
Pico 2
0.08
50.78
9.43
Pico 3
0.08
50.78
8.94
Pico 4
0.08
48.25
9.06
51.41
9.12
Device
Average (Pico)
Nano 1
1
53.54
8.94
Nano 2
1
49.11
9.78
51.32
8.52
Average (Nano)
10"
180
51.84
8.02
10"
180
52.89
7.51
52.42
7.70
Average (10”)
Table III: Log reduction value of bacteriophage φX174 with Sartobind pico and nano devices. MV is membrane volume.
Load volume
(MV)
Pico 1
Pico 2
Pico 3
Pico 4
Nano 1
Nano 2
100
5.4
5.1
5.1
5.3
5.2
5.5
150
5.5
4.9
4.8
5.1
5.3
5.3
Average
5.4
5.0
5.0
5.2
5.3
5.4
ity is very high, exceeding 10 kg of protein
feedstock per liter of membrane volume and
can thus present the risk of membrane fouling. To assess fouling as a function of load
capacity, the Sartobind pico and Sartobind
nano devices were loaded with a 20 g/L
γ-globulin solution to a load capacity of 20
kg/L at a constant inlet pressure of 3 bar. As
seen in Figure 4, while slightly higher flow
decay was observed with the pico device,
the overall flow decay was minimal with
the two devices thus demonstrating the
absence of significant membrane fouling at
high load density.
Characterization of membrane
adsorber devices using model systems
Chromatography media are usually characterized using model molec ules, w ith
dynamic binding capacity and impurity
clearance reported at specific process conditions. The dynamic binding capacity for
Sartobind STIC-PA was determined using
bovine serum albumin (BSA) and DNA, and
impurity clearance was evaluated using
DNA, endotoxin, and bacteriophage.
Dynamic binding capacity: The dynamic
binding capacit y at 10% breakthrough
was measured for the Sartobind pico, the
Sartobind nano, and the Sartobind SingleSep
10” capsule using BSA and DNA model systems. All devices were assembled with
STIC-PA membranes. The breakthrough
curves for the three devices are shown in
Figures 5 and 6 for BSA and DNA, respectively.
The breakthrough curves are similar for all
devices suggesting consistent flow distribution and efficient utilization of the membrane
binding sites at the three scales. Table II shows
the average BSA and DNA dynamic binding
capacity values for several Sartobind pico,
nano and 10” devices. At 10% breakthrough,
the difference in dynamic binding capacity
for all three devices was insignificant. The
consistent dynamic binding capacity with
BSA and DNA supports a linear scalability
from 0.08 mL axial flow pico device to 180
mL radial flow SingleSep 10” capsule.
25
Membrane Scale-Down
Table IV: Endotoxin removal (log reduction value) at pH 7.3 in buffer containing 150 mM NaCl with Sartobind pico and nano
devices. MV is membrane volume.
Load volume
(MV)
Pico 1
Pico 2
Pico 3
Pico 4
Nano 1
Nano 2
50
> 3.96
> 3.96
> 2.92
> 3.96
> 3.96
> 3.96
150
> 3.96
> 3.96
> 3.96
> 3.96
> 3.96
> 3.96
150
> 3.96
> 3.96
> 3.96
> 3.96
> 3.96
> 3.96
Figure 6: DNA breakthrough curves for pico, nano, and 10” devices. MV
is membrane volume.
1
C/C0
0.8
0.6
0.4
0.2
0
0
20
40
60
80
100
Load Volume [MV]
Pico 1
Pico 2
Nano 1
Nano 2
10” capsule 1
10” capsule 2
Removal of bacteriophage: Pathogen clearance was evaluated using the bacteriophage
ΦX174, serving as a surrogate for mouse minute virus (MMV), which is typically used as a
model virus for virus validation studies. Both
ΦX174 (26-33 nm diameter) and MMV (20
nm diameter) are small nonenveloped DNA
viruses with an isoelectric point of around
6.7–7.0 and 6.2 respectively (13). At pH > 7,
both ΦX174 and MMV are mainly negatively
charged and expected to bind to positively
charged AEX chromatography membranes,
resulting in their clearance from protein feedstock through electrostatic interactions. To
compare clearance between Sartobind pico
and Sartobind nano, the same ratio of ΦX174
to membrane volume was loaded. Processscale capsules were not tested because of the
large amount of phage material required. Two
flow-through fractions were collected with
each pico and nano device, and the LRV was
evaluated by comparing the phage titers of
the fractions with the load solution. As shown
in Table III, similar LRVs were obtained at a
load of 100 and 150 MV of phage-spiked buffer, demonstrating linear scalability between
the devices.
26
Removal of endotoxin: Endotoxins are lipopolysaccharides found in the outer membrane
of various gram negative bacteria, can be present as different forms of micelles and vesicles,
and are generally strongly negatively charged.
Because of their ability to elicit immunogenic responses in humans, endotoxins must
be removed to typically < 0.25 Endotoxin
Units per milliliter (EU/mL) where EU is the
unit of measurement for endotoxin activity (USP <29>). Table IV shows the results for
endotoxin removal with Sartobind pico and
nano devices at pH 7.3 in a buffer containing
150 mM sodium chloride. The concentration of endotoxin in the load was 108 EU/
mL, which is significantly higher than the
concentration of endotoxin typically found
in any in-process pools. Three fractions were
collected from the flow-through at loading
volumes of 50, 100, and 150 MV. All flowthrough fractions had an endotoxin concentration below the detection limit of 0.012 EU/
mL resulting in a LRV > 3.96 except one fraction at 50 MV with the pico device. However,
subsequent fractions at higher load volumes
with the same pico device provided LRV >
3.96 which suggests that the anomalous
reading at 50 MV was likely due to an assay
error or sample contamination. Based on
the load volumes tested, the total amount of
endotoxin removal was > 1296 EU with the
pico and > 16200 EU with the nano device.
Significantly larger amount of endotoxin
would be required in the load to saturate the
membrane with the endotoxin molecules to
determine and compare the breakthrough
curves for both pico and nano devices.
Performance of Sartobind pico with
an industrially relevant mAb feedstream
In a mAb purification process, AEX chromatog raphy is t y pically operated in a
flow-through mode to bind trace levels of
impurities such as DNA, putative viruses,
Membrane Scale-Down
Figure 7: Chinese hamster ovary proteins (CHOP) breakthrough curves for
Sartobind pico and nano with a mAb feedstream. MAb feedstock contained 100
ppm CHOP. Experiments were performed at pH 8.0 and 7.0 at 11 mS/cm and at a
flow rate of 10 MV/min.
80
CHOP (ppm)
60
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40
20
0
0
2
4
6
Load density [kg/L]
8
pH 7, 11 mS/cm Pico
pH 7, 11 mS/cm Nano
pH 8, 11 mS/cm Pico
pH 8, 11 mS/cm Nano
endotoxins, and host cell pro teins, wh ile t he m Ab produc t
flows through. The load capacity
is indicated as the mass of product loaded per unit volume of
chromatography membrane (kg
mAb/L membrane) such that the
purity level in the product pool
is acceptable. To assess the performance with an industrially relevant feedstream, both pico and
nano devices were loaded with an
in-process mAb pool post Protein
A and cation exchange chromato g r aphy s te p. S u b s e qu e nt l y,
CHOP levels were monitored in
the f low-through as a function
of load density. The devices were
loaded to 10 kg/L load densit y
at two different solution conditions (pH 7.0 and 8.0 at 11 mS/
cm). CHOP clearance as a function
of load density is shown in Figure
7. Comparable CHOP clearance
was obtained with the pico and
the nano device at both solution
conditions using an industrially
relevant mAb feedstock, suggesting that the Sartobind pico is scalable to the Sartobind nano device.
Additionally, at pH 7.0 and 11
10
mS/cm, a load capacity \ 10 kg/L
could be achieved with pool CHOP
levels < 10 ppm.
The CHOP clearance results are
consistent with the earlier data where
comparable BSA and DNA dynamic
binding capacity was observed
between the pico, nano, and process
scale 10“ devices. Comparable clearance of endotoxin and the bacteriophage further demonstrated the
scalability of Sartobind pico to the
Sartobind nano.
CONCLUSION
It is well documented in the literature that AEX membrane adsorbers are an attractive alternative
to columns for polishing applications in a flow-through mode.
Because of its hydrodynamic benefits, load capacity greater than
10 k g / L of me mbra ne c a n b e
achieved with membrane chromatography. Such high load densit y necessitates a significantly
la rge a mou nt of protei n feedstock for process development
and validation, which could be
cost proh ibit ive. To overcome
this limitation and also to reduce
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27
Membrane Scale-Down
validation cost particularly for virus spiking studies, an ultra scale-down device,
Sartobind pico, having a membrane volume of 0.0 8 mL was developed. Using
mo de l mole c u le s a nd a n i ndu st r ia l ly
relevant mAb feedstock, Sartobind pico
was compared to the existing commercial scale- down dev ice Sartobind nano.
BSA and DNA breakthrough curves, CHOP,
b a c te r io p h a ge, a nd e nd o t ox i n c le a rance data demonstrate the scalability of
Sartobind pico to the Sartobind nano. The
new scale-down pico device will facilitate
the development of flow-through polishing applications for recombinant proteins
and monoclonal antibodies by reducing
the sample consumption by 10 -fold and
providing substantial cost savings for process characterization and virus validation
studies.
REFERENCES
1. J.X. Zhou and T. Tressel, Biotechnol. Prog. 22,
341–349 (2006).
2. J.X. Zhou et al., J. Chromatogr. A 2006,1134,
66–73.
3. J. Glynn et al., “Downstream Procecessing
2010” supplement to Biopharm Int. 22, s16–
s20 (2009).
4. A. Arunakumari, J. Wang, and G. Ferreira,
“Advances in Process Chromatography”
supplement to Biopharm Int. 22 s36–s40 (2007).
5. A. Arunakumari, J. Wang, and G. Ferreira,
“Downstream Procecessing 2010” supplement
to Biopharm Int. 22 s22–s26 (2009).
6. A. Mehta et al., “SBE Supplement–
Bioprocessing” supplement to Chemical
Engineering Progress 104, 14–20 (2008).
7. N. Fraud, Bioprocessing J. 7, 34–37 (2008).
8. A. Pastor, M. Hirai, and S. FischerFruehholz, presentation at GVC/Dechema,
(Osnabrück, Germany, May, 2007).
9. U. Gottschalk, Pharma Focus Asia 7, 60–65
(2008).
10. J. Zhou et al., Biotechnol. Bioeng, 100, 488–
496 (2008).
11. Sartorius Stedim Biotech GMbH, “Scale up
with Sartobind SingleSep,” Application Note,
SL-4042-e07081.
12. R. Faber, Y. Yang, and U. Gottschalk, Biopharm
Int. 22 (10), 11–14 (2009).
13. D. M. Strauss et al., Biotechnol. Bioeng. 104,
371–380 (2009). ◆
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Container Closures
Closures for
Pharmaceutical Preparations:
A Review of Design and
Test Considerations
Design Pics/Kelly Redinger/Getty
ABSTRACT
Closures that form part of the container-closure system are an important component in
the packaging of sterile products. Container-closures maintain the sterility of parenteral
pharmaceuticals and prevent ingress of contamination when a needle is inserted into a
vial. This article describes important aspects to consider in the manufacture of closures
for pharmaceutical preparations, as well as the various physical, chemical, and biological
assessments required to ensure that these closures are fit for purpose.
P
Tim Sandle, PhD,
Bio Products Laboratory, Dagger Lane, Elstree,
WD6 3BX, United Kingdom,
[email protected]
PEER-REVIEWED
Article submitted: June 29, 2012.
Article accepted: July 10, 2012.
32
a rentera l produc ts a re
desig ned, for mu lated, a nd
packaged to be sterile and
to maintain sterility. One of
the most important parts of the packaging of sterile drug products is the
container-closure mechanism. This
article examines the use of closures, for
products intended for injection, in the
pharmaceutical industry. The article
considers the most important aspects
relating to the manufacture of closures
and the different physical and biological assessments required to ensure that
the closures are “fit for purpose.” The
article does not address caps or other
types of seals.
Closures form part of the “containerclosure system.” Container-closures
function to keep the contents of pharmaceutical preparations sterile (e.g., by
providing a barrier between the neck
of a vial and the vial contents) and to
prevent ingress of contamination into
a vial once a needle is inserted (e.g.,
by enabling resealing of the vial after
the needle is withdrawn). The closure,
together with a crimp that creates the
container-closure, and the vial itself
form the primary packaging or packag-
ing component (i.e., the material that
first envelops the product and holds
it) (1). The ideal container-closure will
have low permeability to air and moisture and a high resistance to aging (2).
Therefore, the manufacturers and
users must have confidence in the
quality control and validation of closures. It is an important part of pharmaceutical manufacturing that all
information on the composition and
manufacturing processes for each component type must be understood.
CLOSURES
Pharmaceutical closures, also known
as stoppers or bungs, are an important
part of the final packaging of pharmaceutical preparations, particularly
those that are intended to be sterile.
The most commonly used type of
stopper is the elastomeric closure. An
elastomer is any material that is able
to resume its original shape when a
deforming force is removed, which is
known as viscoelasticity (3).
For the manufacturing of closures,
the elastomer is either natural or, as
is more common, a synthetic rubber,
such as butyl rubber or chlorobutyl
Container Closures
rubber. The advantage of synthetic rubbers
is that the materials are strongly resistant to
permeation by oxygen or to water vapor (4).
In terms of the specification for closures
and the testing and sterilization requirements, the following documents are useful as
starting points:
t '%" Guidance for Industr y: Container
Closure Systems for Packaging Drugs and
Biologics3PDLWJMMF.%.BZ
t &VSPQFBO$PNNJTTJPOGuideline on Plastic
Immediate Packaging Materials (Brussels,
May 2005).
t USP, General Chapter <381> Elastomeric
Closure for Injections.
t E u ro p e a n Ph a r m a co p e i a . C h ap te r 3,
Materials for Containers and Containers.
5IF '%" Code of Federal Regulations (CFR)
QBSU TUJQVMBUFT UIBU DPOUBJOFSDMPsures must provide adequate protection to
the product over the product shelf-life.
Before using a closure in a vial or bottle
with a drug product, the closure must be
assessed to determine if it is suitable for use
with the product that will be filled into the
glass container. The pharmaceutical manufacturer should consider the following questions relating to product compatibility, in
conjunction with the manufacturer of the
closure:
t *TUIFQSPEVDUBCTPSCFECZUIFSVCCFS t %PFT UIF SVCCFS SFBDU XJUI UIF QSPEVDU
BOEMFBDIPVUJNQVSJUJFT t "U XIJDI UFNQFSBUVSF SBOHF JT CPUI DMPTVSFBOEQSPEVDUTUBCMF t )PXFGGFDUJWFJTUIFTFBMJOUFHSJUZ t 8IBUIBQQFOTXIFOUIFQSPEVDUBOETUPQQFSBSFTUPSFEUPHFUIFSPWFSUJNF
Once these questions have been satisfactorily answered, the pharmaceutical manufacturer can work with the manufacturer of the
closure to design the optimal closure for the
vial type and product.
MANUFACTURING PROCESS
The manufacturing process for closures
involves processing raw materials and auxiliary substances; weighing and mixing; followed by vulcanization. Vulcanization is a
chemical process for converting rubber or
related polymers into more durable materials
via the addition of sulfur (or another equivalent curative) together with an accelerating
agent such as 2-mercaptobenzothiazole; an
activator, usually zinc oxide; fillers such as
carbon black or limestone; antioxidants; and
lubricants. Following vulcanization, molding
and compressing occur.
There are two types of molding: compression and injection, of which the former is
the most common. Compression molding is
a method of molding in which the molding
material, generally preheated, is first placed
in an open, heated mold cavity. The mold is
closed with a top force or plug member, and
pressure is applied to force the material into
contact with all mold areas, while heat and
pressure are maintained until the molding
material has cured. Injection molding is a
manufacturing process for producing parts
from both thermoplastic and thermosetting
plastic materials. Material is fed into a heated
barrel, mixed, and forced into a mold cavity
where it cools and hardens to the configuration of the mold cavity.
After molding, the stages are: coating,
washing, siliconization (if required, using
specific, high-viscosity silicon oil), and
packaging. Siliconization has several advantages in that it prevents stoppers from sticking together or onto other surfaces and can
assist with the insertion of a needle through
the stopper. The siliconization step is, however, a potential source of contamination.
Silicone used in the preparation of rubber
stoppers should meet appropriate quality
control criteria and not have an adverse
effect on the safety, quality, or purity of the
drug product.
The mixing of raw materials and auxiliary
substances involves the formulation of the
stopper. A stopper is typically made up of
60% rubber, 30% fillers (which protect the
physical properties of the rubber) and pigments, 5% plasticizers (which provide flexibility), 5% additional chemicals including
accelerators (which help to create the crosslinkages which give the stopper its strength
and hardness), activators (which are a function of the efficiency of the cross-linkages),
and antioxidants (which help to avoid the
degradation of the rubber).
There are different types of rubber, such
as natural rubber (latex), isoprene rubber
(a chemical copy of natural rubber), styrolbutadine rubber, ethylene propylene dyes
monomers, silicone (polysiloxane) rubber,
and halogenized butyl rubber.
33
Container Closures
QUALITY CONTROL OF CLOSURES
A number of quality control checks are
required for the manufacture and release of
closures. These checks include:
After the material has been mixed
t 4QFDJGJD HSBWJUZ XIJDI JT UIF SBUJP PG UIF
weight of the molded piece to the weight
of an equal volume of water
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color chart
t %JTQFSTJPO PG B WVMDBOJ[FE TBNQMF JO SFMBtion to particle size
t &YBNJOBUJPO PG UIF BTI BGUFS CVSOJOH JO
comparison to a reference sample
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Units or an equivalent standard
t 3IFPMPHZ PG UIF DPNQPVOE CZ DPOEVDUJOH
an examination of solids under conditions in
which they respond with plastic flow rather
than deforming elastically in response to an
applied force. This is an assessment of the
force necessary to rotate the material by 1°.
An important distinction is that different materials—types of rubber and formulations—have different profile and respond in
different ways.
Post-compression and molding
The material is checked for rubber thickness
and evenness.
Washing process
t %VSJOH UIF MPBEJOH PG TUPQQFST JOUP B
washer, the quality of the water should
be checked for bioburden and endotoxin
using compendial methods.
t 5IF MPBEJOH PG UIF XBTIFS TIPVME UBLF
place in a controlled environment, usually a cleanroom, with staff appropriately
gowned. Unusually for a pharmaceutical
process and in keeping with cleanrooms
used in the electronics industry, the cleanroom may be fitted with deionization
equipment in order to avoid fibers being
attracted to the rubber.
Post-washing and post-siliconization
After the stoppers have been washed, a number
of quality control checks should be performed.
Mechanical and material tests
t $PNQSFTTJPO UFTU " DPNQSFTTJPO UFTU JT
performed to determine the behavior of
34
materials subjected to compressive loads.
Loading is usually done at a uniform rate
(in/min).
t )BSEOFTT
t 'SBHNFOUBUJPO
t 1FOFUSBCJMJUZ JF XIBU IBQQFOT XIFO
a needle passes through the stopper):
Assessments can be made of the insertion force, break loose force, and extrusion
force. One common issue that can arise is
the generation of rubber particles cut from
the closures when needles are inserted, a
phenomenon sometimes referred to as coring (5).
t "TTFTTNFOUPGEJNFOTJPOTBOEGMFYJCJMJUZ
Many material tests are conducted by testing a selection of closures using a high-speed
color sensor that examines the top, bottomside surface, and inside of the closure.
Physical tests
t 3FTJTUBODF UP TUFSJMJ[BUJPO 5IJT SFRVJSFT
consideration of two questions: how does
the rubber of the stopper react to different
types of sterilization, such as gamma irradiation, ethylene oxide, and steam sterilization, and does the stopper become more
CSJUUMFPWFSUJNF
t 1BSUJDMFUFTUJOH
Chemical tests
t 5FTUT GPS FYUSBDUBCMFT BOE MFBDIBCMFT
& YUSBDUBCMFT BSF DIFNJDBM TVCTUBODFT
that are obtained by exposing the packag ing to a var iet y of solvents under
exaggerated incubation conditions of
time and temperature (6). Leachables
differ from extractables in that they are
chemical substances that migrate under
normal conditions of use from the stopper into a drug product. Leachables are,
therefore, a subset of extractables; all
extractables are potential leachables of
toxicological concern (7).
t 4JMJDPOF PJM EFUFSNJOBUJPO 5IF FGGFDU PG
subvisible silicone particles should be
assessed, because these can cause aggregation with proteins, and the new complex
can potentially trigger an immunochemical reaction within the body of the patient
receiving the drug.
Biological tests
t $ZUPMPHJDBMUFTUJOH
Container Closures
t #JPCVSEFO BTTFTTNFOU TVDI BT <5 CFU/
stopper). Some manufacturers undertake
an examination for mesophilic counts
while others focus on examining for thermophilic bacteria, because such microorganisms will be the most resistant to the
sterilization step.
t #BDUFSJBM FOEPUPYJO UFTUJOH TVDI BT <1
&6TUPQQFS
5IF UFTUJOH PG DMPTVSFT GPS
endotoxin, using the Limulus amebocyte
lysate (LAL) method, is quite difficult in
terms of method validation because the
endotoxin challenge to the rubber surface
can prove to be tricky to recover.
Container-seal tests
Of the different test methods described, the
assessment of the container-closure is arguably the most important because it indicates
whether the device is at risk from extraneous
microbial contamination. Pharmaceutical
containers constructed of materials such as
plastic and glass must be qualified and meet
USP <661> Containers and <671> ContainersPermeation standards. The user will therefore need to undertake additional tests that
examine the physical seal of the closure
in the vial, i.e., when the stopper is fully
inserted and crimped, usually by of an aluminium band. The choice to conduct a physical test or a microbial ingress test for this
purpose is a matter of debate. Some practitioners argue that the physical methods of measuring the system’s integrity are preferred
because they are more reproducible, faster,
less expensive, more reliable, and quantitative. Others argue that, as the objective is to
ensure that the product is safe from microbial contamination, a microbial test is the
only true test. Some opt to undertake both
physical and microbial tests.
A review of industry practices suggests that
failures occur with container-closure seals for
a variety of reasons (8). These failures include
poor quality starting materials, an improper
fit of the container-closure combination, the
lack of sufficient inspection as part of batch
release, insufficient process monitoring or
process control, the use of unreliable manual
or visual inspection techniques, the use of
methods that produce subjective results, and
the lack of proper process validation. The
latter point is addressed through the tests
described below.
Physical tests include the dye test, vacuum
testing, gas leakage determined using a bubble test, liquid leakage detected by atomic
absorption of a copper ion tracer solution,
PS B IFMJVN MFBL SBUF UFTU 0G UIFTF UIF
helium leak test is one of the most widely
conducted; the objective is to detect leaks by
monitoring changes in headspace gas composition or changes in total headspace pressure. This test measures the rate of helium
leak from the vial as well as the actual percentage of helium that is filled within the
vial. Mass spectrometry can be used to measure the rate of leakage. Mass spectrometry-based leak detection is accomplished by
measuring the amount of a tracer gas that
escapes from the container-closure system.
Tracer egress is facilitated by a pressure difference across the container-closure barrier.
Alternative and novel test methods to
assess container-closure integrity include
the use of hygroscopic powder and nearinfrared (NIR) spectroscopy as a means of
visualization. A second example is with airborne ultrasonic technology where a sound
wave is directed towards the container-closure and visualized through the creation of
a high-resolution image. An alternative to
ultrasound is the use of a laser diode or the
utilization of high- voltage technology. These
new techniques have the advantage of being
non-destructive and they allow for a larger
proportion of the batch to be tested, which
increases the level of confidence in the integrity of the seal. These techniques are also
more accurate in allowing identification of
small pinholes, micro cracks and seal imperfections that cannot visually be seen.
8JUI NJDSPCJPMPHJDBM UFTUJOH B TUFSJMity test of the end product or a microbial
ingress test can be considered. The sterility
test is unsuitable because the test will only
detect viable microorganisms present at the
time of the test and those that are capable
of growth within the culture media used.
The microbial ingress test involves direct
microbial challenge and is, therefore, a more
robust test. The objective is to detect microbial ingress based on 1) the probability that
the challenge microorganisms can find a
container-closure leak, 2) the ability of the
microorganisms to traverse the leak, and 3)
the capability of the microorganisms to grow
in the internal container environment.
35
Container Closures
The microbial ingress test can be performed in different ways. One of the key
criteria is the selection of the microorganisms. It is more common to use two different
microorganisms of different sizes and with
different methods of motility. For example,
Brevundimonas diminuta, a very small bacterium, and Escherichia coli, a bacterium with a
relatively powerful motility, are often used in
combination (10). The complexity with the
test relates to achieving a sufficiently high
microbial population.
To conduct a microbial challenge test,
vials are filled with a microbiological growth
medium before stoppering and crimping,
and are immersed in a 35 °C bath containing
magnesium ion as well as 8 to 10 logs of viable bacterial cells for 24 hours. The test units
are then incubated at 35 °C for 7 or 14 days.
Microbial ingress is detected by turbidity and
plating on blood agar.
The described tests, or a selection thereof,
should ensure that the integrity is verified
over the product’s shelf-life, simulating the
stresses the product will be subjected to,
including sterilization, handling, and storage
conditions. The tests, therefore, need to be
made more rigorous in order to simulate “real
life” events, for example by exposing test vials
to stresses of temperature and pressure conditions, which the vials are subjected to when
being transported for distribution and sales.
The level of confidence is increased if three
different batches are assessed. Another option
is to assess vials as part of a stability trial program, which includes a time point at the end
of the shelf-life.
Packaging
After packaging, a selection of bags should
be examined for tears as a part of the quality control assessment. The placement of the
stoppers into the packaging should be underUBLFO XJUIJO BO *40 $MBTT &6 (.1 (SBEF
C cleanroom for standard stoppers and in an
*40 $MBTT &6 (.1 (SBEF " FOWJSPONFOU
for ready-to-sterilize or ready-to-use stoppers.
Sterilization
Closures are typically sterilized by one of
two methods: steam sterilization using autoclaves and gamma irradiation. It should be
noted that not all types of stoppers can be
sterilized by gamma irradiation because the
36
rubber of the stopper will become brittle
from the generation of free radicals in the
polymeric materials (11).
The sterilization of stoppers also requires
the sterilization device to be subject to the
standard tests including thermometric studies and biological indicators for steam sterilization devices and dosimeters for gamma
irradiation.
CONCLUSION
The container-closure system is an essential par t of the f inal presentation of a
pharmaceutical product. It defines the closure, protection, and functionality of a
container while ensuring the safety and
quality of the drug product over the product shelf life. This article has addressed
the important considerations for closures:
the “rubber” stoppers inserted into vials
of products and sealed in place. The article has focused upon the important tests,
control measures, and essential aspects
for ensuring that the product, in its final
packaging, is fit-for-purpose prior to the
administration of the drug. REFERENCES
1. L. Solomun et al., J. Pharm. Biomed. Anal. 48 (3),
744–8 (2008).
2. W. Curry et al., AAPS PharmSciTech. 11 (4), 1572–9
(2010).
3. USP 32–NF 27 (Rockville, MD, 2009), pp. 4133–
4140 .
4. E.J. Smith and R.J. Nash, “Elastomeric Closures
for Parenterals,” in Pharmaceutical Dosage Forms:
Parenteral Medications, K.E. Avis, H.A. Lieberman, L.
Lachman Eds. (Marcel Dekker, New York, 1st ed.,
1992), pp. 445–507.
5. G.D. Chanana, X. Guo, K.E. Avis, et al., J. Parenter.
Sci. Technol. 47 (1), 22–5 (1993).
6. I. Markovic, Am. Pharm. Rev. 9 (6), 20-27 (2006).
7. D.R. Jenke, PDA J. Pharm. Sci. Technol. 59 (4), 265–
281 (2005)
8. FDA Container and Closure Integrity Testing in Lieu
of Sterility Testing as a Component of the Stability
Protocol for Sterile Products, (Rockville, MD, Feb.
2008).
9. L.S. Burrell, M.W. Carver, G.E. DeMuth, and W.J.
Lambert, PDA J, Pharm. Sci. Technol. 54 (6), 449–55
(2000).
10. E. Kirsch et al., PDA J, Pharm. Sci. Technol. 51 (5),
195–202 (1997).
11. P. Kiang et al., J. Parenter. Sci. Technol. 46 (S2)
(1992). ◆
JANUARY 28th - 30th 2013
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www.biomansummit.com
North America’s Leading Biomanufacturing Event Uncovers Tomorrow’s
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Topic and Speaker Highlights Include:
Developing a World-Class Quality
Monitoring System
Ronald C. Branning,
Chief Quality Officer
Genzyme USA
Generating Competitive Advantage and
Improved Performance through Lean
Biomanufacturing Programming
Divakar Ramakrishnan,
VP, Manufacturing Elanco Animal Health
Eli Lilly and Co. USA
Analyzing the Risk of Supply Chain Sourcing
and Manufacturing in Emerging Markets
Andy Skibo,
EVP Operations, MedImmune
Facilities of the Future - What is the Future
of Biomanufacturing?
Jim Miller,
VP and Head, Global Biologic Drug Substance
Manufacturing - Genentech USA
Understanding the Challenges Facing
US Competiveness and Innovation in Bio
Industries
Charles Laranjeira,
SVP, Technical Operations
Cubist Pharmaceuticals Inc. USA
Register now at www.biomansummit.com/1995
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For more information or to confirm
your participation, please contact
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Researched and Produced by:
Operations Survey
Survey: Optimizing Global
Biopharmaceutical Operations
Through Risk Mitigation and Management
Phil Kaminsky, Jiyang Liu,
and Julia Olsen-Claire
A UC Berkeley
survey provides
insight into
biopharma’s
risk concerns
and strategies.
I
ncreasing global competition and
heightened c ustomer e x pec tations have for many years now
encouraged enterprises in a variet y of industr ies to foc us on, and
invest in, effective operations management. In most industries, managers
no longer have to be convinced of the
value of taking an integrated view of
the design, analysis, and operation
of their manufacturing, service, and
logistics operations. Indeed, in many
industries, operational excellence and
a sophisticated approach to supplychain risk management based on flexibility, efficiency, and advanced tools
for logistics network optimization are
no longer a competitive advantage—
they are necessary to compete.
BACKGROUND
Phil Kaminsky, Jiyang Liu,
and Julia Olsen-Claire
work at the CELDi Biopharmaceutical
Operations Initiative at the
University of California, Berkeley.
38
Consider, for example, the semiconductor industry. Thirty years ago, the
semiconductor industry was a growing
technology-focused industry and, for
the first time, was beginning to face
cost pressures. For years, the indust r y had foc used on super ior technolog y, and manufacturing was an
afterthought. As long as the products
were manufactured as they were envisioned by their inventors, there was
little need to pay attention to capacity utilization, operational efficiency,
inventory levels, or risk management;
if you made them, profits would come.
Howeve r, a s t he se m iconduc tor
indust r y mat ured a nd compet it ive
pressures grew, firms began to focus
on operat ions, ut i l i z i ng resou rces
effectively and efficiently, optimizing systems, a nd perhaps most
importantly, dealing effectively with
u nce r t a i nt y a nd r i sk . Sig n i f ic a nt
advances in the science of operations
were required to bring about these
changes, and by working collaboratively among themselves as well as
with academia through organizations
such as SEMATECH, semiconductor
firms were able to make great strides,
pushing the state-of-the-art in semiconductor operations to new heights.
Biopharmaceutical firms now find
themselves in a similar position. As
the biopharmaceutical industry enters
its fourth decade, it is entering a new,
more mature phase. Revenue is growing, innovative business models and
partnerships are being implemented,
and products are coming to market.
At the same time, this new maturity
brings “adult-sized responsibility” (1).
Although the science of biotechnology
is advancing rapidly, with the promise of making an enormous impact
on soc iet y, t he operat ions, supply
chain, and log istics of biotechnology is not keeping pace. The ability
of the industry to reach its potential
requires systems that can produce and
deliver products safely, reliably, and
cost effectively to patients, while also
allowing biopharmaceutical firms to
successfully navigate the many risks
inherent in the industry. It is becoming apparent that the biopharmaceutical supply chain presents a unique set
of operational challenges—demand is
highly uncertain and dependent on
Operations Survey
the results of clinical trials and
competitors’ actions; supply is
highly uncertain; biological processes are complex and incompletely understood; reg ulator y
demands are significant and vary
from region to region, including the existence of agencies in
multiple jurisdictions that add
layers of complexit y; contamination is difficult to detect and
can have a significant impact;
product failures can cost lives;
IP concerns are significant; and
capacity is extremely expensive
and requires long lead times to
build or acquire.
RISK SURVEY
Adding to the complexity, mechanisms for drug production are
typically not standardized even
within firms, technologies cont i nue to c ha nge, a nd gener ic
drugs are poised to dramatically
impact the industry (2). In recent
years, this changing dy na mic
has been a key focus of research
at the University of California,
Berkeley, culminating with the
recent establishment of the CELDi
Biopha r maceut ica l Operat ions
Initiative (BOI), focusing on the
deve lopme nt of c ut t i ng- e dge
tools, techniques, and approaches
to improve production systems,
logistics systems, supply chain,
inventory, and distribution within
biopharmaceutical firms—essentially, biopharmaceutical operat ions. T he resea rc h i n it iat ive
is jointly sponsored by member
firms and the National Science
F o u nd at io n ( N S F ) u nd e r t he
Industry/University Cooperative
Research Program.
Key to the development of this
initiative was a series of industry–academia workshops held at
Berkeley (one sponsored by the NSF)
in which the challenges and opportunities in biopharmaceutical operations management were explored.
Across the industry, managers have
Figure 1: Respondents indicate their level of concern about key risk categories,
where the scale ranges from 1 to 5, with 1 meaning not concerned, 3 meaning
concerned, 5 meaning extremely concerned.
Raw materials
supply risks
Manufacturing
supply reliability
Contamination
Outsourcing-related risk
Forecast errors
Catastrophic
natural events
Other
0
0.5
1
1.5
2
2.5
3
3.5
Figure 2: Respondents, as a percentage, indidcate how they make decisions regarding
inventory level and/or the number of suppliers used (multiple selections were allowed).
Network simulation tools
Inventory/supply chain
optimization tools
Subject matter
experts meeting
Ad-hoc (no system looking
specifically at this)
Other (please specify)
0
an overlapping set of concerns,
and are eager for better approaches,
tools, and techniques that account
for the unique characteristics of biopharmaceutical operations and help
deal with these concerns. A common theme emerged from these
workshops: the need for more effective risk-management tools and
approaches. Many firms are specifically focusing on identifying and
20%
40%
60%
hedging risks associated with their
operations, and are eager to collaborate to improve tools, techniques,
and approaches to do so. As a precursor to a concerted research effort
in this area, the BOI surveyed nearly
300 industry members to explore
attitudes about risk related to suppliers, raw materials, contamination,
outsourcing, disposable technology, demand forecasting, inventory,
39
Operations Survey
Figure 3: Respondents indicate their level of concern about certain manufacturingrelated risks, where the scale ranges from 1 to 5, with 1 meaning not concerned, 3
meaning concerned, 5 meaning extremely concerned.
Contamination
Yield variability
Increasing titers
Disposables
Human resources
and training
Other
0
1
2
3
4
Figure 4: Respondents rank how frequent key risks become an issue.
45.4%
10.1%
42%
Every few years
Once or twice a year
About once a month
More than once a month
40.3%
lighted their concern that firms
in the industry lack a “global” or
“system” view of risks faced, and
rather than developing a cohesive
strateg y to minimize risk, consider risks one at a time, thereby
ignoring their interactions.
Surprisingly given these concerns, however, there is relatively
little focus in the industry on
detailed analysis of relevant data,
relatively little formal quantification of risk, little formal modeling and simulation of risk or risk
mitigation strategies, little focus on
inventory optimization, and little
measurement of uncertainty.
In contrast, in many industries
risk mitigation involves spending
considerable resources collecting
and analyzing data, assessing the
types of variability in the data,
optimizing resource utilization
to mitigate risk, and developing
rigorous models to understand
where inventory and other buffers can most effectively be util i z e d t o h e d ge a g a i n s t r i s k .
Surprisingly, for such a sophisticated industry in so many ways
(e.g., compared with basic cons u me r pro duc t s or i ndu st r ia l
equipment manufacturers), this
industry has a qualitative view of
risk management. This finding is
particularly surprising given the
vast amount of data that is collected in the industry.
WHO TOOK THE SURVEY?
and distribution, with a particular
focusing on understanding which
concerns are most significant, how
firms measure these risks, and what
mitigation strategies they currently
have in place (see sidebar, “Survey
Respondents”). Below is a summary
of the survey’s key observations.
SURVEY RESULTS
Overall, firms are most concerned
w ith qualit y risks, contamina-
40
tion risks, and risks associated
with lack of visibility into contract manufacturing operations.
More broad ly, f i r m s a re concerned with a broad spectrum of
risk-related issues, including supplier risk, manufacturing reliability, inventory risks, cold-chain
issues, a nd forecast ing-related
risks. In preliminary interviews
t hat accompa n ied t he su r vey,
several respondents even high-
The BOI survey was distributed
using a ma iling list compiled
at industry meetings and from
Berkeley’s BOI industry partners.
Although the number of usable
responses varies from question
to que st ion, s u r vey s we r e at
least partially completed. Fortysix percent of respondents work
at biotech/ biopha r m f i r ms,
and an additional 24% work at
pharmaceutical firms. The bulk
of t he rema ining respondents
ident i f y t hemselves as equ ip -
Operations Survey
me nt or r aw-mate r ia l s uppl iers, or as employees of contract
organizations. Twent y-five percent of t he respondents work
at large companies (more than
50,000 employees), 20% work at
very small firms (less than 100
employees), and the rest are dist r ib ute d s ome whe r e b e t we e n
those two extremes. Slightly less
than one quarter of respondents’
firms have no products on the
market, while slightly less than
half have 20 or more products on
the market. In terms of annual
s a le s, re sp onde nt s a l s o come
from a broad spectrum of companies—35% are from large companies with $5 billion or more
in annual sales, and the rest of
t he respondents a re relat ively
equally distributed over the range
from no annual sales to $5 billion in annual sales. Respondents’
firms operate globally, with sales
and manufacturing distributed
around the world, and R&D most
often concentrated on the US East
and West Coasts, and in Western
Europe.
About 30% of survey respondents work i n R& D, a nd 2 0 %
work in manufacturing; the rest
are scat tered throughout var ious roles in engineering, quality,
planning, and so for th. About
half are managers or directors;
the rest are either consultants,
or in various roles ranging from
t he C - su ite to i ndep e nde nt
contractors. Note that the BOI
team made no attempt to limit
responses from a particular firm,
so t hat t he resu lts ref lec t t he
opinions of multiple respondents
within a firm.
OVERALL VIEW OF RISK
O vera ll, respondents a re most
concer ned w it h t he r isk s su rrounding manufacturing
reliability and production cont a m i nat ion, fol lowe d by rawmaterial supply and outsourcing
Figure 5: Respondents indicate their level of concern about certain contractmanufacturing risks, where the scale ranges from 1 to 5, with 1 meaning not
concerned, 3 meaning concerned, and 5 meaning extremely concerned.
On-time delivery/
Reliability
Supplier outages
Visibility into
the process
Quality issues
Ensuring compliance with
regulatory agencies
Other
0
risk. Forecast errors, catastrophic
natural events, regulatory risk,
a nd I P t he f t r a n ke d f u r t he r
behind, although all are of concern to respondents (see Figure
1). Interestingly, given risk concer ns and the inherent uncertainty in this industry, relatively
few respondents rely on quantitative modeling tools to assess
risk: 72% or respondents firms
u s e fa i lu r e mo de a nd e f fe c t s
a n a ly s i s ( F M E A) of r i sk , b ut
fewe r t h a n 3 0 % u s e d i s c r e te
event modeling, and fewer than
27% use Monte– Carlo analysis,
tools that are standard in other
industries. Indeed, while 70% of
respondents monitor batches lost
and on-time performance, half
or fewer mon itor sa fet y stock
levels or customer-service level,
although these are key performance indicators that are useful for measuring the cost and
effectiveness of risk mitigation
strategies.
Raw-material risk
Of all of the raw-material related
risks, single-sourcing stands as
the most significant, followed by
1
2
3
4
contamination (other concerns
included availabilit y, comparabi l it y ac ross vendors, e xt rac tables and leachables, and price
f luctuations). These results are
in spite of the fact that that only
37% of respondents indicate that
their firms use multiple suppliers or similar strategies to mitigate t he r i sk , a lt houg h s ome
responding firms are beginning
to identify diversified sourcing
as a key goa l i n t he produc tdevelopment stage. In addition
to dual sourcing, firms turn to
vendor audits, quality management systems, and large safety
stock s of mater ia l i n order to
mitigate these risks. One responde nt t a l ke d of aba ndon i ng a
“just-i n-t i me” st rateg y due to
the associated risk. Not surprisingly, inventory costs and inventor y levels are the key metrics
t racked to assess raw-mater ia l
acquisition performance (by 58%
and 54% of respondents, respectively). Surprisingly, fewer than
15% of respondents e xpl ic it ly
mention track ing qualit y metrics in this context. In addition,
about ha lf of t he respondents
41
Operations Survey
Figure 6: Respondents indicate how well they explicitly model demand variability
in their planning processes.
44.9%
18.0%
73.0%
We don’t
We model some upsides
and downsides
We use distributions to model
percentiles of likelihood
More than once a month
conce r n s, re sp onde nt s rep or t
primarily turning to high-temperature/short-time (HTST) pasteurization, physical segregation,
and better assay technologies at
roughly equivalent levels (each by
between 45% and 47% of respondents). About 60% report experiencing a contamination event
once or more a year, and roughly
14% experience such an event
once a month or more frequently
(see Figure 4).
29.8%
ment ion foc using on t rack ing
t he f rac t ion of single-sou rced
raw materials. When determining the appropriate inventor ymanagement policies, a surprisingly large fraction of respondents (40%) of respondents do
not use quantitative or scientific
approaches. Slightly over half rely
on optimization tools to make
these decisions, and about 15%
(possibly overlapping with the
optimization users) also use simulation tools (see Figure 2).
Manufacturing-related risk
The most significant manufacturing-related risk as identified
by respondents was contamination, followed in decreasing order
by human resources issues, and
y ield va r iabi l it y. Respondents
were also concerned with increasing titers, but seemed less concer ned w it h t he c ha l lenges
posed by disposables (see section
below). Respondents also identified a variety of other concerns
surrounding product validation,
equipment reliability, and managing capacit y. Sec uring sufficient supply and was also noted
a mong t he respondents’ comments (see Figure 3).
Fou r pr i ma r y st rateg ies a re
42
used to mitigate risks including
training, statistical analysis, quality management techniques, and
a focus on improving both internal and external communication.
About 40% of respondents identify training and education as a
key ma nufac t ur ing r isk-reduction tool, while more than 30%
identify statistical analysis and
quality management. Moreover,
re sp onde nt s rep e ate d ly h ig hlight the positive impact of good
communications both internally
and with CMOs and suppliers on
manufacturing yields.
When addressed specifically,
ma nu fac t u r ing contaminat ion
was reported to be a primary concern of 64% of the respondents
(see next section).
Contamination events
Of the various issues and concer ns sur rounding contamination, media contamination is by
far the largest concern: 39% of
respondents ident if ied t his as
their most significant contamination-related concern. Crossbatc h cont a m i nat ion wa s a
distant second at 19% (followed
by lack of detectability of cont a m i n at ion, a nd e x t r ac t able s
and leachables). To address these
Contract manufacturing
A l m o s t 6 0 % o f r e s p o n d e nt s
report using CMOs for some or all
of their manufacturing. Although
respondents show concern about
all aspects of their relationships
with CMOs (e.g., reliability, visibility, flexibility, and IP) the most
dominant concerns are around
quality and compliance (these are
the only categories for which a
plurality of respondents indicated
“extreme concern”). To address
these risks, the most commonly
noted approaches include a comprehensive focus on prescreening
and monitoring CMOs, frequent
audits, and keeping a representative on the CMO site (and duals ou r c i n g whe r e p o s sible a nd
reasonable) (see Figure 5).
Disposables
Over half of respondents (53%)
currently use disposable technology for commercial manufacturing, and a total of 83% either
plan to or are likely to use disposable commercial manufacturing technology in the future. The
most reported significant risks
around this technology focus, in
decreasing order of reported significance are: comparabilit y as
products are scaled up for commercial runs, timely availability
of supplies from suppliers, and
extractables and leachables. To
add ress t hese r isk s, respondents turn to many of the same
Operations Survey
approac hes t hat t hey u se for
ma nag i ng raw-mate r ia l r i sk s,
includ ing using mu lt iple sup pliers where possible, stringent
vendor qualification and audits,
extensive qualit y control testing and validation, and carefully
constructed contracts specifying
minimum inventory levels.
Forecasting and demand variability
About half (49%) of the respondents consider forecast errors a
significant risk; less than a quarter (24%) are not concerned. In
spite of this, 30% do not explicit ly mo de l de ma nd va r i abi lit y in their planning process,
and another 45% only model some
upside and downside scenarios;
only 18% explicitly model demand
distributions (see Figure 6).
By far, the dominant approach
(70% of the respondents) to dealing with demand uncertainty is
to hold extra inventory—most of
the other cited approaches have
to do with securing extra capacity, either internally or through
relationships with CMOs. About
one third of respondents explicitly
track forecast accuracy by comparing point forecasts with actual
demand, and about one quarter
suggest that their firms do not
explicitly track forecast accuracy.
Distribution
Of the various risks associated
with finished-goods distribution,
cold storage and expiration-related
issues are more of a concern than
contamination, theft, and counterfeiting. No single approach
stands out in the reported mitigation methods for these concerns,
although respondents seem to
continually explore alternative
suppliers and methods for packaging, shipping, and storage (in particular, they mention continual
quality auditing of transportation
service providers, secure packag-
Survey respondents
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ing, tracking systems, diversification, inventor y management
software, and so forth). More than
80% of responding firms store
inventory in multiple sites, selecting these sites in most cases for
their proximity to manufacturing sites or their low operating
costs. Once finished goods shift
to distributor control, only 25%
of respondents are not concerned
with distributors’ risks, although
only 20% are confident in their
visibility into distributors’ risks,
suggesting a key area of potential
improvement.
CONCLUSION AND NEXT STEPS
This sur vey prov ides init ia l i nsights i nto t he c u r rent
state of risk management and
r i s k- m it i g at io n s t r at e g i e s i n
t he biopha r maceut ica l i ndus try. In subsequent surveys, the
BOI tea m i ntend s to e x plore
s p e c i f ic r i s k- m it i g at ion ap p roaches and metrics c urrently
employed by progressive firms, in
order to develop a better understanding of which metrics, tools,
and approaches are particularly
useful. At the same time, there is
clearly a need within the biopharmaceutical industry for analytical
tools and approaches that account
for the specific characteristics of
this industry, and that use the
vast quantities of data available to
help managers make more rigorous, informed, model-based decisions to manage and mitigate the
complex set of risks faced by the
industry.
REFERENCES
1. Ernst & Young, “Beyond Borders: Global
Biotechnology Report,” 2007.
2. R. Johnson and P. Kaminsky,
“Biopharmaceutical Operations:
Developing the Science,” PharmaFocus
Asia 9 (2008). z
43
SGS Life Science Services
Company Description
SGS Life Science Services is a leading
contract service organization providing
clinical research services,
analytical development,
biologics characterization
a n d q u a l it y c o nt r o l
testing. SGS provides
clinical trial management
(Pha se I to I V ) a nd
ser vices encompassing
bioa na ly t ic a l te st i ng ,
data management,
biostatistics, and
regulatory consultancy.
SGS also offers contract
analytical services (detailed below) that
include analytical chemistry, microbiology,
stability studies, method development, and
protein analysis. With the recent acquisition
of Vitrology, SGS expands its service offering
for biologics in the areas of cell bank and
virus seeds characterization, raw material and
bulk harvest testing (sterility, mycoplasma,
viruses), final product testing for residual
DNA, and safety consultancy services. SGS
is the world’s leading inspection, verification,
testing, and certification company.
SGS LIFE SCIENCE SERVICES
75 Passaic Ave.
Fairfield, NJ 07004
TELEPHONE
866.747.5003
FAX
973.244.1823
EMAIL
[email protected]
WEBSITE
www.sgs.com/lifescience
NUMBER OF EMPLOYEES
Life Science Services
worldwide: 1500
SGS Group: 70,000
YEAR FOUNDED
1878
44
Technical Services
t Quality control testing of raw materials,
APIs, and finished products
t Monograph testing (USP, EP, BP, and JP)
t A nalytical method development and
validation
t Antibody product analysis
t Bioana lysis, mass spectrometr y and
immunoassays
t Biologics safety testing (endotoxin, virus
and mycoplasma)
t Cell line characterization
t Container testing (extractables and
leachables)
t Environmental Monitoring
t Glycosylation Analysis
t
t
t
t
t
t
t
t
Host cell impurity testing
Virus testing (characterization of Cell Banks)
Molecular biology services
Medical device testing
Microbiological testing
Protein/peptide analysis and quantification
Safety testing (in vitro)
Stabilit y testing according to ICH
guidelines or customer specifications
t Water system validation
t Clinical trials from Phase I-IV
t Data management and statistics
Facilities
With truly global coverage and a strong
local footprint, in North America, SGS
has facilities in Lincolnshire (Illinois),
Fa irf ield (New Jersey), West Chester
(Pennsylvania), and Mississauga (Canada), as
well as a clinical trial management office in
Gaithersburg (Maryland). SGS’s laboratories
operate according to high quality standards
(cGMP, GLP, ISO 17025) and have been
inspected by FDA or local regulator y
authorities.
Markets Served
SGS ser ves various life science
companies, including pharmaceutica l,
biopharmaceutical, biotechnolog y, and
medical device manufacturers. SGS operates
a global network of 19 Life Science Services
laboratories with facilities in the US, UK,
C a nad a, Belg iu m, Fra nc e, Germa ny,
India, China, Taiwan, Singapore, and
Switzerland. The Top 20 pharmaceutical
and biopharmaceutical companies trust SGS
as a partner for their quality control testing
needs.
ADVERTORIAL
THE LARGEST NETWORK OF
CONTRACT ANALYTICAL LABS
SGS LIFE SCIENCE SERVICES
SGS Life Science Services is a leading contract service organization providing analytical development, biologics characterization,
biosafety testing, quality control product release, as well as Phase I-IV clinical research services. Whether your organization
is large and global or small and regional, rely on SGS as your partner for outsourced testing.
Operating a harmonized network of 19 laboratories in 11 countries, SGS offers lean quality standards, reliability and
regulatory / technical expertise... all within close proximity to you.
© 2012 SGS Société Générale de Surveillance SA – All rights reserved
CONTACT US:
Europe: +32 10 42 11 11
North America : +1 866 747 5003
Asia : +65 63790 111
[email protected]
SGS IS THE WORLD’S LEADING INSPECTION, VERIFICATION, TESTING AND CERTIFICATION COMPANY
WWW.SGS.COM/PHARMAQC
WHEN YOU NEED TO BE SURE
Global Markets
Navigating Emerging Markets
In an introduction
to a new
BioPharm
International
series on
manufacturing
within global
markets,
the author
provides key
considerations for
getting started.
Jill E. Sackman, DVM, PhD,
is a senior consultant at
Numerof & Associates, Inc. (NAI), St. Louis,
MO, www.nai-consulting.com.
46
T
he global market for pharmaceuticals continues to grow rapidly,
and in response, manufacturers
are moving to align their market strategy and product pipeline to meet
emerging, unmet needs. Companies
exploring opportunities in global markets
face dynamic demographic and disease
trends, changing market demands, and
evolving regulatory requirements—all of
which differ from one country to another.
This complex environment makes planning for global market entry into both
developed and developing countries a
moving target.
In the face of such complexity, organizations need to be prepared to navigate
constantly changing, inconsistent, and
diverging regulatory and market trends
in a structured, but flexible manner that
minimizes rework and maximizes value.
Whether the goal is to outsource the manufacture of US products to India to lower
costs, or to market imported or domestically developed products in China, a consistent approach for understanding market
trends and navigating foreign regulation of
pharmaceuticals is crucial for developing a
successful global market strategy. With that
Chad Baker/Getty Images
Jill E. Sackman
said, it is important to allow for variability
in regional requirements (sometimes even
within a single country), market demands,
and product pipelines.
Over the next year, I’ll be tackling some
of the region-specific challenges that
pharmaceutical companies face in a new
BioPharm International column. This introductory article provides some key considerations for navigating and managing diverse
market and regulatory trends across the
globe in a consistent manner.
DEVELOP A COMMON APPROACH
TO NAVIGATING REQUIREMENTS
The globalization of pharmaceutical markets has fueled an emphasis on improving
the regulation of these industries across
developed and developing countries.
Ideally, steps towards harmonization would
be among the goals of improving regulation. Countries are instead taking independent strides at varying paces to improve
the regulation of markets that are very different from each other. Across the board,
submission requirements vary and enforcement resources are limited, which can contribute to frustrating waiting periods and
prolonged product-approval timelines.
Global Markets
This degree of regulatory variability may seem daunting, but approaching regional requirements in a consistent manner can make the process more manageable.
Although regulatory oversight and enforcement varies
from country to country, there are common regulatory
steps that play an integral role in shaping each stage of
the product development, approval, manufacturing, and
marketing lifecycle. Regardless of the region or product
in question, the regulatory process will likely involve the
following: product classification, preclinical testing, clinical
evaluation, product registration, manufacturing/production
approval, quality system management, and compliance
with good manufacturing practice (GMP) as well as good
clinical practice (GCP), and good laboratory practice (GLP).
At each step in the regulatory process, authorities may
establish approval requirements, regionally accepted best
practices, recognized credentials and certifications, and/
or quality system requirements. The controls that are put
in place at each step in the regulatory process can—and
will—differ from country to country, and from product
to product. Being able to identify and understand these
controls for any given country or region will be crucial
because they may have implications for timelines, processes, cost, and revenues.
Moreover, understanding the nature and stringency of
these regulatory measures will be important. These factors
may impact how effectively a product can be brought to
market in other regions, under a completely different set
of regulatory requirements.
evolved over time may help organizations prepare to navigate trends in a rapidly changing region. As an example,
China and Brazil are developing health technology infrastructures along the lines of the UK’s National Institute for
Health and Clinical Excellence (NICE). Understanding what
that process has looked like could be helpful in preparing
for future regulatory demands in these developing markets.
DETERMINE THE MOST EFFICIENT
ALLOCATION OF RESOURCES
As part of developing a consistent approach, it will be
important to determine the most efficient way to identify
and collect information and to manage your organization’s
ongoing presence in foreign markets. Consider centralizing
the allocation of resources, as appropriate, to minimize
duplication of effort to understand regulatory and market
trends for a particular region. By centralizing these efforts,
you can ensure that one division of your organization is
not building networks or partnerships in regions where
you already have those relationships developed.
To ensure that your company is able to navigate foreign
market trends and changing regulatory requirements on
Continued on p. 57
GAIN INSIGHT INTO
REGIONAL MARKET TRENDS
The implications of population growth, demographic
changes, and shifting lifestyles on disease trends and market demands differ tremendously from region to region.
As part of determining market potential within a given
region, it will be important to understand these trends
within the context of your technical requirements (e.g.,
product specifications, materials.) and operational needs.
Additionally, regional health policy and reimbursement
trends will have a tremendous impact on successful market entry, and should serve as criteria for developing your
market strategy. Specifically, it will be important to understand in-region: access to healthcare, epidemiological
trends, current clinical treatment paradigms (care paths),
market demand (i.e., taking into consideration the patient
population as well as clinical insight from in-region providers), competitive set/competing products, reimbursement for similar products , and data requirements (i.e.,
clinical and health economics) for market access.
It is also important that the decisions your organization makes as part of its global strategy account not only
for current trends, but also for the expected or anticipated
future environment. Looking at how similar markets have
47
Boot Camp: Tech Guide
Glycan Analysis: A Primer
I
n this fourth part of a series of primers with
training experts from the National Institute
for Bioprocessing Research and Training
(NIBRT), Pauline Rudd, PhD, professor of glycobiology at University College Dublin (UCD), discusses glycan analysis. NIBRT provides training,
educational, and research solutions for the international bioprocessing industry in state-of-theart facilities. Located in South Dublin, it is based
on an innovative collaboration between UCD,
Trinity College Dublin, Dublin City University,
and the Institute of Technology Sligo.
KEY DEVELOPMENT AND
MANUFACTURING CONSIDERATIONS
BioPharm: Can you provide a brief overview of what
exactly glycan analysis targets and its importance
in bioprocessing?
Rudd: Most glycoproteins, and almost all of
the new biological drugs, are proteins with sugars
attached to them. These sugars are important for
the safety and efficacy of drugs, so it is necessary
to be able to control the processing of the sugar
structures to make sure that the drug is as effective
as possible. There are many aspects of developing
and processing a drug that require having analytical technologies for glycosylation. For example, in
the beginning of a drug-development process,
one needs to understand the role of the sugars
on the protein being used. On erythropoietin,
for example, the sugars must be multiantennary
and fully capped with sialic acid; otherwise, the
erythropoietin will only be in the patient for a few
minutes, whereas if it is completely sialylated, it
will be present in the patient for 3 hours or more,
during which time it will be able to be effective
in stimulating the production of red blood cells. It
is key to monitor the production process to make
sure that the erythropoietin has the sugar structures that provide the full benefits of glycosylation.
It is always important for biologics developers to
understand exactly how the sugars in their product
are going to modulate the functions of the drug in
48
the patient. Once this knowledge is obained, drug
developers can define an optimal glycan profile.
The next stage of bioprocessing, clonal selection,
requires identifying a high-producing clone that
has the ability to fully glycosylate the molecules
with the optimal sugars. When selecting clones,
glycan analysis enables the producer to determine
the complement of glycoenzymes that are operating within a particular clone.
The next aspect of glycosylation that must be
checked is the potential introduction of an antigenic epitope. If one is making a product in a
nonhuman cell line, such as mouse cells, it is necessary to check the levels of alpha-galactose and
N-glycolyl-neuraminic acid residues, which may
be antigenic to humans. After the candidate clones
are selected, the process of producing the protein
from the cells must be monitored. Taking samples
during the process allows one to assess whether
the media composition is optimal for producing
the desired glycosylation profile. In process development, optimizing the media is necessary to
produce high levels of correctly folded proteins as
well as the desired post-translational modifications,
including glycosylation.
BioPharm: What role do glycans play in the
manufacturing stage?
Rudd: When making a biologic product, one
needs to track the glycans, which will help to
determine the best time to harvest the product.
When one reaches the downstream processing
stage, high-performance liquid chromatography
(HPLC) is often used to select subfractions of
the product. Being able to analyze the glycans to
ensure that their subfractions are correctly glycosylated is crucial. This information will be used
during conversations with regulatory authorities
so that the agency can agree on specifications
for the drug product. Glycan analysis will be
part of that specification because it will be part
of the regulators assessment of the drug’s safety
and efficacy. It will also be necessary to demonstrate to regulators that the process is robust and
December 2012
Sveta Demidoff/Getty Images
NIBRT’s Pauline Rudd on what to expect when performing glycan analysis.
Leveraging the Unique Attributes
of Chromatography Resins to Drive
Productivity and Decrease Cost of Goods
0N-DEMAND WEBCAST
Register free atwww.biopharminternational.com/resins
EVENT OVERVIEW
In order to address current industry manufacturing challenges
and demands, process development scientists must balance
a number of factors when designing a downstream process
including capacity, resolution, process flexibility, disposability,
throughput and cost. Increased titers and product demand
have caused a substantial bottleneck in downstream processing
for both antibodies as well as recombinant proteins. There is a
greater need to purify molecules more efficiently and to reduce
the costs associated with purification.
This webcast will discuss
New solutions on the chromatography side of purification to
improve process performance and increase process flexibility
How high performance chromatography can impact
biotherapeutic cost of goods
Examples of how to generate more productive and cost
effective processes to address downstream challenges
Who Should Attend
Biopharmaceutical companies, biotech companies, and contract
manufacturing organizations with titles that include:
Process Development
Process Engineering
Quality Assurance/Control/Validation
Key Learning Objectives
Discuss new approaches for the
downstream purification processes to
obtain improved impurity clearance
and maximize yields
Discuss how to obtain improved
purity and throughput, while
increasing process flexibility within a
purification scheme
Discuss how chromatography can
benefit downstream processing and
impact cost of goods and overall
productivity
Speakers
Shelly Cote Parra
Sr. Applications Scientist
Life Technologies
Christine Gebski
Head of the POROS® Resin
Business and Global Applications
Life Technologies
Moderator
Angie Drakulich
Editorial Director
Anything to do with manufacturing
Presented by
Or, any customer interested in the manufacturing/production of
biomolecules such as monoclonal antibodies, therapeutic proteins,
and vaccines.
Sponsored by
For questions, contact Sara Barschdorf
at [email protected]
can be reproduced for batch and lot
release.
Then comes the consideration
of long-term storage. Although in
general, glycans are quite stable,
it is important to ensure that glycans do not change by testing after
degradation and stability studies.
Functional assays are also important
to determine whether a product, IgG,
for example, is able to bind to the
desired receptor and not to those
receptors that can cause side effects
in the patient. Some of these questions require an understanding of
how glycosylation modulates activity
of the drug.
Overall, there are many reasons to
perform glycan analysis and many
ways to approach it. It is therefore
important to understand the question being asked before deciding
which method to use.
COMMON CHALLENGES IN
CONTROL AND TECHNIQUE
BioPharm: W hat common challenges is the industry facing today
when performing glycan analysis?
Rudd: Glycan processing is difficult to control because it involves
a complex process that involves
the expression of genes (the genes
carry the code for the glycosylating enzymes) as well as the delivery
of monosaccharides on nucleotide
donors to grow the glycans. There are
many things to control, and nearly
600 proteins are required to build
the glycans as well as to transport the
glycoproteins into the correct organelle for complete glycan processing.
If one is over-expressing a protein, it
is possible to exhaust some of the glycosylation machinery. It is not uncommon to find incomplete structures
attached to a protein, which indicate
that the processes to build the sugars
has not operated on all copies of the
glycoprotein going through the secretory pathway. Understanding how to
get the cells to express at a level where
the rest of the machinery can cope is
just one challenge.
50
Another difficulty is determining
what to do when the glycosylation
is going wrong. One needs to understand in depth how media components and metal ions, for example,
can alter glycan analysis. The natural
cell is exquisitely tuned and responds
very finely to its environment. It is
difficult to replicate this robustness
in a bioprocesssor.
BioPharm: You mentioned that
industry uses various techniques
for glycan analysis. What are the
most commonly used instruments
and techniques?
Rudd: Glycan analysis depends
on a series of separations technologies that exploit different physical
properties of the oligosaccharides.
There are several ways to approach
it. If you are focused on analyzing
released glycans, you need to have
an optimized method for releasing
them. There are many glycoconjugates, but from the point of view of
the pharmaceutical companies the
most commons ones are N-linked
glycans. There is an enzyme called
PNGase F that can be used to remove
them from proteins of most species. After that stage, various separations technologies can be used to
separate glycans according to mass/
charge, charge, size, and lipophilicity
by techniques such as hydrophilic
interaction chromatography (HILIC),
ion-exchange chromatography,
reverse-phase chromatography, or
capillary electrophoresis.
BioPharm: What gaps still exist in
glycan-related technology and instrumentation?
Rudd: NIBRT uses a lot of special
instrumentation, but we are working to get the analytical technologies
miniaturized and as straightforward
as we can. We have a robot that
can accommodate 96 well plates,
for example, so that one can put
the samples on the instrument and
come back later to put the released
sugars on a HPLC machine or use
mass spectrometry for analysis. We
also have the capability to run lin-
December 2012
ear samples, meaning that we can
take samples one at a time from a
bioprocesser every few hours. Going
forward, industry will be looking to
miniaturization, automation, and,
particularly, automated data analysis.
REGULATORY ISSUES
BioPharm: What regulatory expecta-
tions exist for glycan analysis when
developing a biologic compared with
a legacy product?
Rudd: Actually, there is a debate at
the moment. Regulators need companies to report critical features of
the glycosylation, but in some cases,
it is not clear what “critical” features
include. There is a need for more
basic research to clarify these questions. For some molecules, such as
IgG, it is known that the Fc glycosylation modulates effector function, so the regulators can ask for a
full glycan analysis of IgG. One can
report the sialylated structures, the
levels of galactosylated, fucosylated,
and bisected structures, because it
is known that each of these features
can modulate a function. To get the
information, teams need to perform
a complete analysis of IgG and present the data in a way that answers
the questions about critical features
that affect the safety and efficacy of
their product.
If one is working with erythropoietin, it is necessary to report the
percentages of different antennary
structures as well as the extent of
sialylation, because this critically
affects the pharmacokinetcs of the
drug. One needs to report levels of
N-glycol-neuraminic acid, alpha(1,3)
linked galactose as well as levels of
xylose and alpha(1-3)-linked fucose,
because these are potential antigenic epitopes. In general, regulatory expectations are getting higher
because the technolog y is getting better. In the past, none of us
really understood the implications
of glycosylation in therapeutics
Continued on p. 56
MAY 20-22, 2013
Sheraton San Diego
Hotel and Marina
San Diego
Call For Papers
Contributed Papers
Submission Deadline:
February 13, 2013
SUBMISSION DETAILS:
www.aaps.org/nationalbiotech
Bioanalytical Best Practices
Bioanalytical Methods for Sample Cleanup
Preparation of biological samples for chromatographic analyses.
52
The choice of sample
preparation method should
depend on the quality of
the data required.
tion method should depend on the quality
of the data required. It makes little sense to
invest weeks of development time attempting
to achieve pg/mL sensitivity for a screening
assay. However, it may be important to invest
such time to develop and validate a method for
a lead drug candidate undergoing human safety
assessments that are subject to FDA regulatory
scrutiny. Indeed, validating analytical procedures is the process of determining a suitable
method that is capable of providing useful analytical data. It is important to bear in mind that
a method that is valid in one situation could be
invalid in another (10, 11).
In the pharmaceutical industry, the most
common biological sample matrix is plasma.
Moreover, it is common practice to dilute troublesome matrices like urine or cerebrospinal
fluid with plasma and apply previously developed plasma extraction protocols. Drugs are
most commonly isolated from plasma using
one (or occasionally, a combination) of either
liquid–liquid extraction, protein precipitation,
or solid phase extraction. Other less common
choices include column-switching (LC–LC),
affinity extraction, and ultrafiltration.
LIQUID-LIQUID EXTRACTION
Liquid-liquid extraction (LLE) using organic
solvents offers sample cleanup with analyte
December 2012
Image: PASIEKA/Science Photo Library/Getty.
H
igh-throughput bioanalytical methods
are essential to support the rapid discovery and development of drugs in
the pharmaceutical industry. Liquid chromatography coupled with tandem mass spectrometric detection (LC–MS/MS) is considered
as the benchmark analytical methodology for
quantifying new chemical entities in biological fluids (1–4). Because of the high sensitivity
and selectivity of LC–MS/MS, the time required
for method development and subsequent sample analysis is dramatically reduced. Rigorous
chromatographic resolution of analytes and/
or tedious sample extraction protocols are typically not required even when complex biological matrices are used. Most chromatographic
techniques have matured and automation is now
commonplace (5–7). Nevertheless, with common
sample analysis times of less than three minutes,
the bottleneck in sample analysis has become
the sample preparation step. Sample preparation
is still considered to be a slow and labor-intensive
process, and it is rare for an analyst to be able to
inject samples directly into an LC–MS/MS system with no pretreatment.
The importance of sample preparation stems
from three major concerns—removing interferences from the biological sample matrices, concentrating the
analyte(s) of interest, and improving
analytical system performance (8).
An industry survey noted a marked
increase in methods requiring limits of quantitation of less than 1
ppb, and the trend toward trace
analyses is not diminishing (9).
Roger N. Hayes is vice-president and Optimized sample preparation techgeneral manager of laboratory sciences niques that provide high enrichat MPI Research, ment factors become crucial for
54943 North Main Street, these dilute concentrations.
Mattawan, MI 49071.
The choice of sample prepara-
enrichment steps, and is a rugged off-line sample preparation
process that is well suited for routine high-t hroughput LC – M S/
MS analysis. The basic concept
of LLE is to partition an analyte
into a volatile organic solvent
away f rom polar proteins and
lipids that remain in an aqueous phase. The organic phase is
removed, evaporated, a nd t he
sample reconstituted for injection
onto an LC –MS/MS system. By
careful choice of organic solvent,
LLE is amenable to automation in
a 96-well format (12–15). In order,
the acceptability of solvents for
automated LLE is methyl t-butyl
ether > 95/5 hexane/ethanol >>
ethyl acetate. LLE does, on occasion, suffer from emulsion formation, which may be resolved by
extended centrifugation.
LLE can also be performed in the
solid state by using diatomaceous
earth (Hydromatrix or Celite).
Several such products are available
commercially for performing supported liquid extractions (SLE) in a
96-well plate format.
PROTEIN PRECIPITATION
Plasma sample preparat ion by
protein precipitation (PPT) is the
most widely used technique for
LC–MS/MS analysis because of its
simplicity, low cost, and universality. PPT is amenable to automation in a 96-well format (16).
Precipitation of plasma proteins
is most commonly performed by
using organic solvents like acetonitrile or methanol. Following
denaturation, the sample is cent r i f uge d a nd t he sup er nat a nt
is d i rec t ly i njec ted. However,
organic solvents are inefficient in
precipitating proteins and often
require signif icant dilution of
the plasma sample, typically by
two- to three-fold. Overcoming
the impact of dilution by injecting larger volumes of the plasma
extract may be precluded when
using reversed-phase HPLC
because of the high organic content. Evaporation of the extracted
samples to near dryness followed
by reconstitution in an appropriate solvent is usually required. In
order, the preference of solvent
for automated PPT is 95/5 (v/v)
acetonitrile/acetone > acetonitrile
>> methanol.
ability, ultimately such a loss in
sensitivity limits the achievable
limit of quantification. 96 -well
filter plates are available commercially and they provide the
analyst the ability to conduct, in
an automated fashion, the protein crash procedure, the mix
step, and the filtration of protein
precipitants in the same 96-well
flow-through plate.
Assessing the potential
for ion suppression
over the elution time
is an important step in
developing a rugged
method.
SOLID-PHASE EXTRACTION
Alternative choices for precipitation of plasma proteins include
trichloroacetic acid (TCA) or zinc
sulfate. Reagents like TCA and
zinc sulfate, however, are unable
to remove small proteins, polypeptides, and salts, thereby contributing to high ionic strength
of the supernatant that can subsequently suppress ionization and
attenuate LC–MS/MS response.
Despite the attractiveness of
PPT as a rapid approach to sample preparation, potential drawbacks exist. For example, because
only proteins are removed, ion
s u p p r e s s io n f r o m c o - e lut i n g
matrix components can significantly reduce sensitivity, especially when using electrospray
ionization. Therefore, assessing
the potential for ion suppression
over the elution time is an important step in developing a rugged
method. Although using stable
isotope-labeled internal standards
can compensate for response vari-
December 2012
The third alternative to sample
cleanup is solid-phase extraction
(SPE). SPE met hods consist of
loading samples onto pre-conditioned sorbent-filled cartridges,
often arranged in a 96-well plate
format. Loaded samples are then
washed with an appropriate solvent, and the analyte(s) is subsequently eluted. Method selection
and extract cleanliness are determined by the retention mechanism and the ability of the wash
stage to effectively remove endogenous components. For example,
ionic interactions, as represented
by cation and anion exchange,
offer a greater degree of analyte
selectivity and, therefore, extract
cleanliness when compared to
C18-based sorbents. In general,
extracts from SPE are cleaner than
those from PPT.
SPE ga i ne d i n p opu la rity because of its compatibility
with automation, especially with
sorb e nt mate r ia l pac ke d i nto
a 96 -well format plate (17–18).
Te c h nolo g ic a l i mp r o v e m e nt s
include the development of polymeric SPE sorbents that no longer
suffer from sorbent drying problems while enjoy ing extended
working pH ranges. Taking advantage of the full pH range of the
sorbent, a specific pH and organic
modulated SPE (i.e., an optimized
SPE) method can be developed
to provide clean sample extracts.
T he hyd roph i l ic-hyd rophobic
nature of these polymeric sup-
53
p or t s i s a me nable to ge ne r ic
extraction techniques.
A ge ne r ic p r o to c ol s ho u ld
achieve high recovery; however,
high recoveries do not necessarily correlate with high sensitivity in LC–MS/MS. Achieving high
sensitivity is usually a trade-off
between recovery and chemical
interference or ion suppression.
Nevertheless, high sensitivity and
high recovery are achievable by
selective retention of a basic drug
using strong cation exchange SPE.
Fortunately, the majority of drug
candidates have a basic functionality that can be leveraged for
selective retention on an appropriate strong cation exchange SPE
material.
A s de s c r ib e d, SPE met ho d s
often involve evaporation and
subsequent reconstitution of the
eluent before LC–MS/MS analysis. T hese steps not only take
time and effort, but can also lead
to the loss of valuable sample.
Therefore, the ability to elute in
ver y small volumes of solvent
is desirable to minimize sample
pr e pa r at ion t i me a nd r e duc e
sample loss. Low sorbent mass
and novel 96-well plate designs,
including SPE pipette tips and
discs, have allev iated some of
these concerns (19).
Another approach to consider
is the direct coupling of SPE to
the LC–MS/MS system (i.e., online SPE) (20). Differences in flow
rates du r ing load a nd elut ion
steps afford additional opportunities to enhance the extraction
process. For example, sufficiently
high flow rates can induce turbulent flow chromatography that
actually involves a combination
of size-exclusion and adsorption
phenomena (21–24). If the analyte
fraction has a high enough affinity for the stationary phase inside
the pores, then it w ill remain
there until a solvent w ith the
54
appropriate strength desorbs it.
Online SPE methods have the
potential to significantly enhance
sensitivity because no dilution of
sample occurs. Eliminating analyte collection, evaporation, reconstitution, and injection not only
improves reproducibility, but also
saves time, labor, and solvents.
CONCLUSION
T he genera l idea of sa mple
cleanup is that all elements of a
method should contribute to its
required sensitivity and selectivity. Issues to consider when selecting a bioanalytical method for
sample cleanup should include
what matrix the analy te is in,
the detection limit and dynamic
range required, the number of
samples to be analyzed, analyte
stability to extraction, and the
amount of matrix available.
The final method should be
orthogonal to maximize selectivity and reduce ion suppression. If
C18 SPE is used for sample extract ion, t hen t he a na lyst should
c on sid e r c at ion e xc h a n ge or
phe nyl colu m n c h romatog ra phy. Alternatively, if strong cation exchange SPE was selected
for sample extraction, then any
reverse-phase LC method can be
used. The sample cleanup method
should be assessed for recovery,
selectivity, precision, accuracy,
and ruggedness. Formal validation may also be required (25–27).
REFERENCES
1. T.R. Covey, E.D. Lee, and J. Henion,
Anal. Chem. 58 (12) 2453–2460
(1986).
2. H. Fouda et al., J. Am. Soc. Mass
Spectrom. 2, 164–167 (1991).
3. E.C. Huang et al., Anal. Chem. 62,
713–725 (1990).
4. R.S. Plumb et al., Xenobiotica, 31
(8–9), 599–617 (2001).
5. D. Mole, R.J. Mason, and R.D.
McDowall, J. Pharm. Biomed. Anal. 11
(3), 183–190 (1993).
December 2012
6. E. Doyle et al., Anal. Proc. 26, 294–
295 (1989).
7. M. Jemal, Biomed. Chromatogr. 14 (6),
422–429 (2000).
8. R.D. McDowall et al., J. Pharm. Biomed.
Anal. 7 (9), 1087–1096 (1989).
9. R.E. Majors, LCGC North America, 20,
1098–1113 (2002).
10. B.A. Persson, J. Vessman, and R.D.
McDowall, LC-GC Int. 11, 160–164
(1998).
11. B.A. Persson, J. Vessman, and R.D.
McDowall, LC-GC Int. 10 (9), 574–576
(1998).
12. N. Zhang, K.L. Hoffman, W. Li, and
D.T. Rossi, J. Pharm. Biomed. Anal. 22,
131–138 (2000).
13. Z. Shen, S. Wang, and R. Bakhtiar,
Rapid Commun. Mass Spectrom. 16
(5), 332–338 (2002).
14. J. Zweigenbaum et al., Anal. Chem. 71
(13), 2294–2300 (1999).
15. M. Jemal, D. Teitz, Z. Ouyang, and S.
Khan, J. Chromatogr. B 732 (2), 501–
508 (1999).
16. D. O’Connor, D.E. Clarke, D. Morrison,
and A.P. Watt, Rapid Commun. Mass
Spectrom., 16 (11), 1065–1071(2002).
17. C. Sottani, C. Minoia, M. D’Incalci,
M. Paganini, and M. Zucchetti, Rapid
Commun. Mass Spectrom. 12 (5) 251–
255 (1998).
18. H. Simpson et al., Rapid Commun.
Mass Spectrom., 12 (2) 75–82 (1998).
19. R.E. Majors. New Designs and Formats
in Solid-Phase Extraction sample
preparation, LCGC Europe 12 (1) 2–6
(2001).
20. F. Beaudry, J.C. Yves Le Blanc,
M. Coutu, and N.K. Brown, Rapid
Commun. Mass Spectrom 12 (17)
1216–1222 (1998).
21. H.M. Quinn and J.J. Takarewski,
“Improvement in Chemical Analyses”
International Patent Number
WO97/16724 (May, 1997).
22. M. Jemal, Zh. Ouyang, B.C. Chen,
and D. Teitz, Rapid Commun. Mass
Spectrom. 13 (11) 1003–1015 (1999).
23. J.T. Wu, H. Zeng, M. Qian, B.L.
Brogdon, and S.E. Unger, Anal. Chem.
72 (1) 61–67 (2000).
24. M. Jemal, Y. Qing, and D.B. Whigan,
Rapid Commun. Mass Spectrom. 12
(19) 1389–1399 (1998).
25. J.R. Kagel, W. Donati, L.E. Elvebak,
and J.A. Jersey, Amer. Lab., 33 (24)
20–23 (2001).
26. V.P. Shah et al., Eur. J. Drug Metab. 16
(4) 249–255 (1991).
27. A.R. Buick, et al., J. Pharm. Biomed.
Anal. 8 (8–12) 629–637 (1990). ◆
Final Word
Patents and
Postapproval Batch Testing
Can postapproval FDA filings immunize
pharma companies from patent lawsuits?
It shall not be an act of infringement to make, use,
offer to sell, or sell . . . a patented invention . . . solely
for uses reasonably related to the development and
submission of information under a Federal law which
regulates the manufacture, use, or sale of drugs or veterinary biologic products (2).
But what happens if the company is sued for
infringement after it obtains approval from
FDA? Until recently, most in the legal industry would have said that postapproval batch
testing is not protected by Hatch–Waxman.
As recent as 2011, in Classen Immunotherapies,
Inc. v. Biogen IDEC, the Federal Circuit Court
of Appeals—the final authority on most patent matters—decided that the safe harbor does
not apply to “information that may be routinely reported to the FDA, long after marketing approval has been obtained” (2). Classen
had accused Biogen and GlaxoSmithK line
(GSK) of infringing Classen’s patent relating
to an immunization method (3). The patented
method involved screening immunization
schedules and selecting and administering the
schedule that presented the lowest risk of developing certain immune-mediated chronic disorders later in life (4). As part of an FDA study,
Biogen and GSK used the patented methods
to provide vaccines, advise on immunization
schedules, and report adverse vaccine effects
to FDA (5). Biogen and GSK contended that
If the Hatch–Waxman Act
protects postapproval batch
testing, the value of patents on
analytical methods, particularly covering biosimilars, may
significantly decrease.
their activity fell within the safe harbor (6).
But the Federal Circuit disagreed, concluding
that the provision only provides a safe harbor
to expedite the development of information for
regulatory approval of generic counterparts of
patented products. GSK and Biogen were held
not immune from Classen’s suit.
But i n Aug u st 2 012 , t he s a me cou r t
decided in Momenta Pharmaceuticals, Inc. v.
Amphastar Pharmaceuticals, Inc., (7) that the
safe harbor provision indeed might immunize
post-approval activities. The patent at issue
in Momenta covered a method for analyzing
batches of enoxaparin, a synthetic version of
the blood-thinning agent heparin (8). Because
of enoxaparin’s unique chemical makeup,
FDA requires batch analysis as a condition
for the post-FDA approval sale of the drug
(9). Momenta claimed that Amphastar’s quality control testing of its enoxoparin batches
i n f r i nged Moment a’s patent; A mphast a r
argued that its testing fell within Hatch–
Waxman’s safe harbor because FDA required
the testing (10). The Federal Circuit determined that § 271(e)(1) unambiguously covers
any activity reasonably related to developing
M. Freeman/PhotoLink/Getty Images
F
DA requires that pharmaceutical companies create and maintain pre-approval
batch records for both generic and brand
drugs. In general, a company can do so without risking infringing any patents covering
manufacture of a drug. Congress enacted the
so-called “safe harbor” provision of the Hatch–
Waxman Act in 1984 specifically allowing
pharmaceutical companies to test their products prior to obtaining regulatory approval to
market them (1). Under that provision:
Kevin Murphy is a partner with the New
York office of Frommer Lawrence and Haug
LLP. Andrew Nason is an associate in
the DC office of the same firm, KMurphy@
flhlaw.com. The views expressed in this
article are solely those of the authors
and are not to be attributed to Frommer
Lawrence and Haug LLP or any of its clients.
55
Final Word
and submitting information under
a federal law that regulates the
manufacture, use, or sale of drugs
regardless of whether the activity
occurs pre- or postapproval (11).
The decision has major consequences for the industry. If the
Hatch–Waxman Act protects postapproval batch testing, the value
of patents on analytical methods,
particularly covering biosimilars,
may significantly decrease.
C h ie f Judge R ade r, who
aut hor e d t he m ajo r it y o p i n ion in Classen, issued a lengthy
and shar ply worded dissent in
Mo m e n t a , c r it ic i z i n g h i s f e l low judges for failing to follow
Classen (12). Judge Moore, who
authored the majority opinion in
Momenta and a dissent in Classen,
saw it differently. She squared
the Classen and Momenta decisions not i ng t hat, i n Classe n,
there was no requirement that
Biogen or GSK submit any data
to FDA. By contrast in Momenta,
FDA required analytical records
for each batch of enoxoparin produced in order to maintain regulatory approval (13).
W het he r one c a n re conc i le
these two decisions, the Supreme
Cour t could ver y well end up
resolv ing t he issue. GSK f iled
a petition for certiorari asking
t he Supreme C ou r t to c la r i f y
“[w]hether the Federal Circuit’s
i nte r p r e t at io n o f § 271(e)(1)
[in Classen], which a rbit ra r ily
restricts the safe harbor to preapproval activities, is faithful to
statutory text that contains no
such limitation, and decisions
of this Court rejecting similar
ef for ts to impose extratext ual
limitations on the statute” (14).
The Supreme Court may welcome
the opport unit y to prov ide its
guidance on the scope of Hatch–
Waxman’s safe harbor—an issue
t he h igh cou r t has add ressed
twice before (15)— especially in
light of this seeming inconsistenc y in Federa l Circ u it precedent in t his a rea of t he law.
REFERENCES
1. Eli Lilly & Co. v. Medtronic, Inc., 496 U.S.
661, 669-71 (1990).
2. 35 U.S.C. § 271(e)(1) (2012).
3. 659 F.3d 1057, 1060, 1070
(Fed. Cir. 2011).
4. Ibid. at 1060.
5. Ibid. at 1070-72.
6. Ibid. at 1070.
7. 686 F.3d 1348 (Fed. Cir. 2012).
8. Ibid. at 1351.
9. Ibid. at 1353.
10. Ibid. at 1352-53.
11. Ibid. at 1354-55.
12. Ibid. at 1361-76.
13. Ibid. at 1357-58.
14. Petition for a Writ of Certiorari at
i., GlaxoSmithKline v. Classen
Immunotherapies, Inc., No. 11-1078
(U.S. Feb. 28, 2012).
15. Merck KGaA v. Integra Lifesciences I,
Ltd., 545 U.S. 193, 206-07 (2005); Eli
Lilly & Co. v. Medtronic, Inc., 496 U.S.
661 (1990). ◆
Continued from p. 50
and regulatory requirements were to
define the glycosylation as well as
one could. Our understanding has
moved a long way since then, and
regulations are far more demanding.
Readers can listen to the full interview
with Dr. Rudd via an audio podcast on
BioPharmInternational.com. ◆
KEY TAKEAWAYS
t 6OEFSTUBOEUIFSPMFPG
TVHBSTPOZPVSQSPUFJO
t #FBXBSFPGWBSJPVT
JOTUSVNFOUBUJPOPQUJPOT
UPQFSGPSNTFQBSBUJPO
BOBMZTFT
t ,FFQVQUPEBUFXJUI
UIFJNQSPWFNFOUTJO
CJPJOGPSNBUJDTGPSEBUB
JOUFSQSFUBUJPO
56
BIOPHARM’S BOOT CAMP GUIDES
Below is a list of all the BioPharm International Boot Camp guides that
exist to date, including articles and podcast interviews. Check them out on
www.BioPharmInternational.com/BasicTraining. If there is a topic we
haven’t covered yet that you would like to see in a future issue, please email
the editor at [email protected].
Business guide topics:
t Outsourcing Strategies t Considerations when Working with Suppliers
t GMP Facilities and Operations Management t The Drug-Development
Game: Risks and Rewards t Product Differentiation t Virtual Biotech Setups
t Industry Training t Commercializing a Compound t Project Management
t Pipeline Growth t Implementing Knowledge Management t Changing Your
Business Model
Technical guide topics:
t Biosimilars Development t Reference-Product Dilemmas t Downstream
Processing t Upstream Processing t Product Characterization t Aseptic
Processing and Fill/Finish
Ad Index
Company
Navigating Emerging Markets –
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Eurofins Lancaster Laboratories
30, 31
Life Technologies
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Nova Biomedical
58, 59
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44, 45
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an ongoing basis, organizations also
frequently decide to pursue partnerships with in-region resources
that are familiar with current and
impending clinical, regulatory,
reimbursement, and competitive
landscapes. Additionally, in-region
resources are prepared to handle
cultural nuances that you may not
anticipate, which may ultimately
have an impact on timelines, costs,
and successful market entry.
Knowing when to outsource is
important, but managing in-region
partnerships closely is equally crucial. Most regions are moving to
regulate in-region firms as an extension of the manufacturer that partnered with them, and therefore,
centralized management of partners is more crucial now than ever
before. A consistent mechanism for
ensuring that you have the necessary auditing capabilities in place
will be key to the success of these
relationships.
CELL LINES
Continued from p. 47
BE PREPARED FOR CHANGE
The demands placed on drug-product companies operating on a global
basis are changing dramatically.
New regulatory requirements have a
significant impact on matters, such
as product design, commercialization, and market-entry strategies.
Additionally, demographic and disease trends, evolving policy changes
and shifting market demands complicate navigating foreign markets in
an efficient manner.
These dynamic, unpredictable,
and region-specific considerations
introduce a great deal of risk for
companies. However, for companies that have developed the capabilities to anticipate and adapt to
change, the current and impending global environment presents tremendous opportunity. ◆
You can hear additional tips from
Dr. Sackman in a podcast interview on
www.BioPharmInternational.com.
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200 Prospect Street
Waltham, MA 02454
TELEPHONE
781.894.0800
Company Description
Nova Biomedical is a technology company based
in Waltham, Massachusetts that provides stateof-the-art automated cell culture analyzers for
the biopharmaceutical market. In 1998, Nova
introduced the first line of chemistry analyzers
for the biotechnology industry—the BioProfile® series.
In 2007, Nova added the
BioProfile® Flex series with
an expanded menu and realtime analysis of key metabolites, nutrients, and gases in
cell culture media. These
fully automated, multi-test
systems simultaneously measure the following parameters: pH, glucose, lactate,
glutamine, glutamate, pO2, pCO2, ammonium,
sodium, potassium, phosphate, IgG, calcium,
chloride, and cell viability and density. In 2011,
we announced the BioProfile CDV Automated
Cell Density/viability analyzer that can measure
up to 80 million cells/mL and provide results in
less than 3 minutes.
BioProfile analyzers are intended to optimize
process development, enhance process reliability and reproducibility, improve manufacturing
yield and quality, and reduce testing time and
cost. In fact, BioProfile analyzers can replace up
to seven different instruments and measurement
techniques while dramatically decreasing space
requirements and capital equipment, labor, and
reagent costs.
An optional On-Line Autosampler allows the
user to connect multiple spinner flasks, bioreactors, and other vessels directly to the BioProfile
analyzer, and establish user-defined sampling
frequencies and feedback instructions for each
reactor vessel. All results are stored in a Data
Management System for subsequent retrieval or
transmission to a data historian.
key nutrients, metabolites, and gases in cell culture and fermentation media. Our all-in-one
BioProfile FLEX analyzer provides immediate
measurement of key chemistries, cell density/
viability, IgG, phosphate, and osmolality in cell
culture media to provide a total picture of cell
growth in a single instrument. The BioProfile
FLEX OPC Connectivity is a complete plug and
play solution that facilitates connectivity with all
OPC-compliant systems. Our optional On-Line
Autosampler connects up to 10 bioreactors to one
BioProfile FLEX, providing real-time testing.
BioProfile Analyzers feature:
t Simultaneous measurement of up to 15 key
parameters
t A full 15 test profile including IgG in eight
minutes
t Automatic one button operation
t Direct sampling from syringes, tubes, flasks,
and pipettes
t 0.5 mL sample size
t No sample dilution or centrifugation
t Automatic analyzer calibration
Cell Density/Viability Analyzer
Nova’s BioProfile CDV incorporates advanced
technology for rapid, accurate, high resolution
measurement of cell density and cell viability at
a moderate cost. Based on the widely recognized
trypan blue exclusion method, BioProfile CDV
combines advanced technology in robotics, optics
and computer algorithms in a highly automated
analyzer.
t Easy to use, intuitive interface
t 12-position sample tray for walk-away sample
analysis
t High resolution vision system
t Advanced computer algorithms for fast image
processing and classification
BioProfile products provide assays necessary to
address the testing requirements of process development, pilot plant, and production facilities.
Developed in conjunction with leading biotechnology companies, these test menus enable
control of the most critical constituents of cell
culture media.
FAX
781.894.5915
EMAIL
[email protected]
WEBSITE
www.novabiomedical.com
58
Major Products
Cell Culture Chemistry Analyzers
BioProfile® series automated chemistry analyzers are designed for real-time, rapid analysis of
ADVERTORIAL
One Automated Analyzer for
Fast Comprehensive Cell Culture Analysis
The Power of One
Flexible Modular Design
BioProfile FLEX can reduce cell culture analysis time, labor, and
operating costs by consolidating multiple analyzers into a single,
easy to use workstation.
BioProfile FLEX can be customized with 1–4 modules to consolidate up
to 15 vital cell culture tests:
Module 1: Glucose, Lactate, Glutamine, Glutamate, Ammonium,
pH, PCO2, PO2, Sodium, Potassium, Calcium
Module 2: Cell Density/Cell Viability
Module 3: Osmolality
Module 4: IgG
One compact workstation, up to 15 Cell Culture assays,
including IgG
One small, 1 mL sample conserves cell culture mass and end product
One fast, 6 minute analysis saves at least one hour over multiple
instrument analyses
One integrated data source simplifies data collection, analysis,
and archiving
One validation saves time vs validation of multiple instruments
New IgG Module
A new IgG measurement combines protein binding methodology with photometric
endpoint detection, to provide:
IgG results in less that 6 minutes
No sample preparation or centrifugation
0.1 to 10.0 g/L measurement range
Other Options
On-Line Autosampler with automated sampling from up to 10 bioreactors
OPC Connectivity automates data and control commands
Test Menu
Gluc
Lac
Gln
Glu NH4+
pH
PO2 PCO2 Na+
K+
ZZZ%LR3URÀOH)/(;FRP
Ca++
CD
CV
Osm
IgG

December 2012

Transcription

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