Document Detail

Rapid production of clinical-grade gammaretroviral vectors in expanded surface roller bottles using a "modified" step-filtration process for clearance of packaging cells.
MedLine Citation:
PMID:  20662590     Owner:  NLM     Status:  MEDLINE    
Production of clinical-grade gammaretroviral vectors for ex vivo gene delivery requires a scalable process that can rapidly generate large amounts of vector supernatant, clear large numbers of residual packaging cells with minimal decreases in vector titer, and satisfy all current regulatory guidelines regarding product biosafety. To that end, we have developed a simplified method that is compliant with current good manufacturing practices for the production of clinical-grade gammaretroviral vectors in a clinical research environment. We validated a large-scale production platform utilizing 1,700-cm(2) expanded surface roller bottles and a "modified" step-filtration process consisting of a 40/150-μm dual-screen filter for aggregate removal followed by a Sepacell 500II leukocyte reduction filter for removal of residual packaging cells. This clarification process can clear at least 2 × 10(9) viable producer cells using a single filter set-up without any significant loss of titer post-filtration. This platform typically generates 18 liters of vector supernatant to support small-scale clinical trials, but can easily be scaled up to 70 liters during a single manufacturing run. To date, this platform has generated five clinical-grade gammaretroviral vector products, four of which are now being used in adoptive cell therapy clinical trials for the treatment of a variety of solid cancers.
Steven A Feldman; Stephanie L Goff; Hui Xu; Mary A Black; James N Kochenderfer; Laura A Johnson; James C Yang; Qiong Wang; Maria R Parkhurst; Scott Cross; Richard A Morgan; Kenneth Cornetta; Steven A Rosenberg
Publication Detail:
Type:  Journal Article     Date:  2010-12-12
Journal Detail:
Title:  Human gene therapy     Volume:  22     ISSN:  1557-7422     ISO Abbreviation:  Hum. Gene Ther.     Publication Date:  2011 Jan 
Date Detail:
Created Date:  2011-01-21     Completed Date:  2011-04-28     Revised Date:  2013-05-29    
Medline Journal Info:
Nlm Unique ID:  9008950     Medline TA:  Hum Gene Ther     Country:  United States    
Other Details:
Languages:  eng     Pagination:  107-15     Citation Subset:  IM    
Surgery Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.
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MeSH Terms
Cell Culture Techniques / methods*
Cell Line
Gammaretrovirus / genetics*
Genetic Therapy
Genetic Vectors*
Virus Cultivation / methods

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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