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