Cincinnati Children`s Hospital Translational Laboratory

T R A N S L AT I O N A L
L A B O R AT O RY
Experimental Hematology
and Cancer Biology
Cincinnati Children’s Hospital
Translational Laboratory
The Cincinnati Children’s Hospital Translational Laboratory
was established in 2003 to efficiently move innovative ideas
from the bench to the bedside in a manner that is both cost
effective and that meets quality assurance requirements. We
offer a one-stop shop to support clinical trials of novel gene
and cell therapies, along with specialized laboratories for
basic research, testing, scale-up, and clinical work.
The Translational Laboratory consists of:
•Vector Production Facility (pharmaceutical grade products)
•Cell Manipulations Laboratory (research and
pharmaceutical grade products)
•Translational Trials Development and Support Lab
(diagnostics, clinical trial support)
•Viral Vector Core (research grade products)
•Development Laboratory
The Lab operates under current Good Manufacturing
Practices (cGMP), Good Laboratory Practices (GLP) and
Good Tissue Practice (GTP) in state-of-the-art facilities.
Clinical grade products are produced in ISO Class 7
cleanrooms designed for aseptic processing, which provide
a safe method for production of pharmacuetical grade
biologics and innovative cellular based therapies suitable
for use in Phase I/II clinical trials. A new cleanroom facility
was opened in 2009 to specifically meet the demanding
needs of the scientific community.
The testing laboratory has been certified by the American
College of Pathology and Clinical Laboratory Improvement
Amendments (CAP/CLIA). In addition to routine diagnostics,
the testing lab also specializes in developing and carrying
out unique esoteric laboratory procedures for use in
support of clinical trials.
We recognize that every project is unique and we work
closely with Sponsors to transition non-clinical bench top
protocols to fully scalable and reproducible GMP/GTP
or CAP/CLIA compliant processes. Our solution-oriented
strategy enables our scientific team to identify and address
technical issues in a time and cost effective manner.
This approach expedites the translation of your process
technology streamlining the move of your project from
bench to clinic.
The leadership of the Translational Lab are active members
of the Association of Academic Biologics Manufacturers
(AABM) and provide leadership in education and regulatory
compliance.
For more information
Punam Malik, MD, Director, Translational Laboratory
phone 513- 636- 8588
email [email protected]
Diana Nordling, Operations Director,
Translational Laboratory
phone 513 - 803 -1063
email [email protected]
web
www.cincinnatichildrens.org/translationalcores
T R A N S L AT I O N A L
L A B O R AT O RY
Viral Vector Core
The Cincinnati Children’s Hospital Medical Center
Viral Vector Core offers an outstanding infrastructure
and personnel for the production of a wide range of
research or GLP quality viral vectors and stable producer
cell lines. The Core also offers non-GMP quality control
testing including vector titer by flow cytometry or
PCR. Other services include the concentration and
purification of vectors using ultra-concentration, ultrafiltration and chromatographic methods.
Research grade vector products:
•Lentiviral vectors
•Gamma retroviral vectors
•Foamy viral vectors
•Adeno-associated viral vectors
•Adenovirus
Non-GMP quality control testing:
•Infectious titer by flow cytometry or qPCR
•Genomic titer by qPCR
•P24 titer by ELISA
•Sterility
Process development and scale-up services for
novel vectors, as well as specialized assays to support
detailed vector characterization, can also be provided.
The Viral Vector Core works closely with the
GMP-compliant Vector Production Facility to
ensure seamless processes and translation
from preclinical studies to human gene therapy
clinical trials. Scientific personnel assist clients
with the development and optimization of novel
vector systems.
The laboratory staff specializes in the scale up,
characterization and technology transfer of preclinical
vector processes to the GMP environment.Since
its establishment in 2004, the Viral Vector Corehas
produced more than five thousand vector products
foracademic, industry, and other clients
For more information
Scott Cross, MS, Manager
phone 513-803-1067
email [email protected]
Han van der Loo, PhD, Director
phone 513-803-1066
email [email protected]
web
www.cincinnatichildrens.org/translationalcores
Experimental Hematology
and Cancer Biology
T R A N S L AT I O N A L
L A B O R AT O RY
Vector Production Facility
The Vector Production Facility, established in 2003, is one
of the pillars of the Translational Laboratory at Cincinnati
Children’s Hospital and is dedicated to the production of
viral vectors in support of early phase clinical trials. The
facility — housed in ISO Class 7 cleanroom production
suites and support rooms —includes research laboratories
for process development and scale-up, along with quality
control, controlled storage rooms for pre- and postproduction products.
The new (2008) state-of-the-art 10,000 ft2 cleanroom
facility is compliant with current Good manufacturing
Practices (cGMP), and includes segregated production
suites, allowing the simultaneous production of clinical
grade products.
Our staff are experienced in the production of gammaRetroviral Vectors, Lentiviral Vectors, Adeno-Associated
Viral Vectors, Adenovirus and Foamy virus vectors, as
well as the production of Master Cell Banks. We have
produced and certified vectors for Phase I/II clinical trials,
with products released in both the US and Europe (UK)
for clients in academia, government and industry. Services
include certification testing, stability studies, purification,
concentration, and vialing, as well as short-term cryogenic
storage of clinical products, validated shipping, and
manufacturing of products for Good Laboratory Practices
(GLP) toxicity studies.
All of our cGMP operations are supported by dedicated
Quality Assurance staff. The facilities and equipment
are fully validated and calibrated to National Institute of
Standards and Technology (NIST) standards. We have a
Type V Drug Master File (MF-BB) on file with the FDA/CBER.
Letters of cross-reference are available upon request.
We work closely with clients to transition research
grade vector production to a fully scalable and
reproducible GMP compliant process. As an
academically-based not-for-profit, we pride ourselves
in delivering the highest level of quality, while staying on
budget and on time. Ultimately, our goal is to improve
outcomes of therapies and to make a difference for patients.
For more information
Han van der Loo, PhD, Director
phone 513- 803 -1066
email [email protected]
web
www.cincinnatichildrens.org/translationalcores
Experimental Hematology
and Cancer Biology
T R A N S L AT I O N A L
L A B O R AT O RY
Experimental Hematology
and Cancer Biology
Pluripotent Stem Cell
Characterization Core
Induced pluripotent stem cells (iPSC) are revolutionizing
research in developmental biology, regenerative medicine,
and cellular disease modeling. A major obstacle for the
development of iPSC based therapies is the high degree
of technical skill and wide range of expertise required
to fully characterize human iPSC lines. Our Core
includes experienced scientists specialized in stem
cell and molecular biology, as well as genomics and
bioinformatics to provide a complete characterization
of human iPSC. These services are managed through the
Translational Laboratory at Cincinnati Children’s Hospital.
We provide a number of iPSC characterization assays,
including:
Standard Assay Panel:
•Morphology
•Flow cytometry for markers of pluripotency and
differentiation
•Immunofluorescence microscopy
•Standard karyotype
•Quantitative RT-PCR 92 gene expression array
(expression of “stemness” and “differentiation”
genes is compared to that of ES cells)
Comprehensive Assay Panel:
•Teratoma formation and basic histologic analysis
•mRNA-seq
•miRNA-seq
•DNA methylation analysis for 450,000 CpG sites
•High resolution karyotype (copy number variation)
•Integration site analysis (for retroviral and
lentiviral vectors)
Differentiation Assay Panel:
•Basic unbiased in vitro embryoid body differentiation
and mRNA-seq
•Targeted germ layer differentiation and mRNA-seq:
– Definitive endoderm
– Mesoderm
– Ectoderm
The Translational Cell Manipulation Lab supports the
process development/scale-up of innovative therapies
for Phase I/II clinical trials. We partner with researchers to
carry new cell-based therapies from laboratory bench to the
clinic by working closely with each Sponsor/PI, to develop
clinical grade cell manipulation/culture process and timeline
that meets program needs and regulatory requirements.
For more information
Carolyn Lutzko, PhD, Co-Director CPML
phone 513- 803-2420
email [email protected]
web
www.cincinnatichildrens.org/translationalcores
T R A N S L AT I O N A L
L A B O R AT O RY
Experimental Hematology
and Cancer Biology
Cell Manipulation Laboratory
The Cell Manipulation Laboratory (CML) is part of the
Translational Laboratory at Cincinnati Children’s Hospital.
We support the development, scale up and processing of
innovative cell-based therapies for Phase I/II clinical studies.
This laboratory ensures reproducible production of large
scale quantities of therapeutic cells under strict quality
control and safety conditions required by the FDA for
human studies.
We partner with researchers to carry potential new
cell-based therapies from laboratory research to clinical
study by working in close contact with each Sponsor/PI,
and developing a culture process and timeline that meets
program needs and regulatory requirements.
Cell services include:
•enrichment or depletion of specific cell subsets
•ex vivo antigen priming
•cytokine and antibody-based activation
•ex vivo genetic modification (transduction)
•expansion of cells
•cryopreservation and/or preparation for infusion
Our expertise is in culturing a wide variety of primary
human cell types including:
•hematopoietic progenitors and stem cells
•mesenchymal stem and progenitor cells
•embryonic and induced pluripotent stem cells
•immune cells including dendritic cells and lymphocyte
sub-populations
•endothelial and epithelial cell
•tumor cells
•hematopoietic populations (erythroid, myeloid and
B- and T-lymphoid populations)
All more-than-minimally manipulated cell and tissue
procedures are performed in aseptic clean rooms in
accordance with GMP and GTP quality compliance
requirements. All procedures utilized by the CML were
developed to meet FACT standards, and labels are designed
to meet the requirements of ISBT-28.
CML staff have experience in providing investigators support
in development of relevant CMC (Chemistry Manufacturing
and Control) sections for the gene transfer for both INDs
and IMPDs.
The CML holds a Type V Master File (MF-BB) with the FDA/
CBER that describes the details of the facility and process
control. A letter of cross reference to the DMF is available
upon request.
For more information
Carolyn Lutzko, PhD, Co-Director CPML
phone 513 - 803-2420
email [email protected]
Diana Nordling, Co-Director CPML
phone 513 - 803 -1063
email [email protected]
web
www.cincinnatichildrens.org/translationalcores
T R A N S L AT I O N A L
L A B O R AT O RY
Experimental Hematology
and Cancer Biology
Clinical Trials Support Laboratory
Accredited by the College of American Pathologists
(CAP) and in compliance with Clinical Laboratory
Improvement Amendments (CLIA) regulations,
the Translational Trials Development and Support
Laboratory (TTDSL) — part of the Translational
Laboratory at Cincinnati Children’s Hospital —
performs diagnostic tests and supports early phase
clinical (gene transfer) trials. Assays are performed per
standard operating procedures with established controls
and can be validated to meet CAP/CLIA requirements.
Molecular and cell-based assays can be tailored to
meet your requirements. Some of our unique
diagnostic tests include:
•Fanconi anemia complementation group analysis
•FancD2 Western blot analysis
•VEGF-D quantitation in human serum (ELISA)
•Clonogenic assays (CFU)
•Endotoxin by LAL
Preclinical and clinical support of gene transfer
trials include:
•Sequence-specific real time PCR
•Insertion site analysis (LM or LAM)
•Development of study specific procedures
•Sample processing
•In vivo murine safety studies
•In vitro immortalization assay (IVIM)
The TTDSL routinely performs preclinical safety
studies in support of IND applications to the FDA
for gene transfer trials that use integrating vectors.
We also provide assay support for cell manipulation
and vector production development, as well as
release testing of vector products and transduced
cells. TTDSL has designated Quality Assurance
personnel to support the Laboratory.
The TTDSL holds a Type V Master File (MF-BB) with
the FDA/CBER that describes the TTDSL’s standard
operating procedures. A letter of cross reference to the
DMF is available upon request.
For more information
Elke Grassman, PhD, Director
phone 513- 636 - 0958
email [email protected]
web
www.cincinnatichildrens.org/translationalcores