For all types of RSAs for laboratory services

• The scientific research in gene therapy that is the subject of the RSA must be consistent with the NHLBI Mission (http://www.nhlbi.nih.gov/about/org/mission.htm) or, if the research addresses a disease area consistent with the mission of another NIH Institute, that Institute must agree to cover the cost of the service.
• The investigator must complete the on-line Registration Form on the GTRP main web page and receive approval by the NHLBI Gene Therapy Group (GTG) allowing the investigator to submit an RSA.
• RSA approval and implementation is contingent upon availability of funds in the GTRP. 
• Services provided through an approved RSA are at no cost to the investigator.
• The various types of RSAs are independent of one another.  An applicant Investigator may submit any type of RSA for a qualified gene therapy research project regardless of past or future utilization of GTRP resources for the project.

In addition, for Clinical-Grade AAV Vector Production

• The applicant must have adequate preclinical data and the requisite regulatory and other oversight reviews for the proposed clinical protocol in which the clinical-grade vector will be used.
• Funding must already be secured and documented for the proposed clinical trial.

A  Request for Service Application (RSA) is the mechanism for applying for GTRP resources. Investigators must register on-line and be approved by the NHLBI Gene Therapy Group in order to access the Request for Service Application.

In order for the laboratory to meet the vector production requirements of the clinical study, the applicant investigator will be asked to provide such information  as:

• A detailed and comprehensive history of the vector (cis) construct, including a copy of the sequencing report of the transgene cassette region (ITR to ITR) for the actual material provided.  The vector plasmid should contain a KanR (not AmpR) antibiotic resistance gene and a plasmid backbone length >6kbp to prevent undesirable “reverse” packaging during clinical vector manufacturing.  These features are required to assure purity and safety of the clinical vector product.
  
  
• A detailed report and standard procedure for quantitative measurement of transgene expression in vitro (e.g. transduction combined with transgene immunoassay or functional assay), including provision of specialized reagents (e.g. cell lines, antibodies) required to perform the procedure.
  
  
• Additional information as required to ensure safety, purity, potency, and stability of the clinical vector product.

The applicant investigator is encouraged to contact the laboratory for assistance in completing the RSA. 

The Clinical Vector Core, a division of The Center for Cellular and Molecular Therapeutics at The Children’s Hospital of Philadelphia is a state-of-the art facility established to support translational/clinical studies using recombinant AAV gene transfer vectors.

Vision: To help realize the promise of AAV gene transfer vectors to address significant unmet medical needs.

The cGMP Facility: The state-of-the art cGMP vector manufacturing suite, established in 2005, boasts dedicated staff with extensive experience in AAV-based clinical vector development and manufacturing.  A highly optimized helper virus-free transfection process used for vector generation/biosynthesis is coupled to an efficient purification process that includes ion exchange chromatography and density-based separations to achieve optimal vector purity and yield.  The processes have been developed for several AAV subtypes/serotypes, and provide a Final Product with minimal levels of process (e.g. residual culture medium and host cell derived) and product-related (e.g. empty AAV capsids) impurities. An extensive series of Quality Control assays are performed on manufacturing process intermediates and on the Final Product to ensure vector product safety, purity, potency, and stability.  Our Quality Assurance program, which includes extensive Process and Environmental Monitoring, provides full documentation of vector quality at all stages of clinical product manufacturing for each Lot prepared.  AAV vectors prepared using these methods demonstrated excellent safety in clinical studies.