Production scale-up and validation of packaging cell clearance of clinical-grade retroviral vector stocks produced in Cell Factories Journal Article


Authors: Przybylowski, M.; Hakakha, A.; Stefanski, J.; Hodges, J.; Sadelain, M.; Riviere, I.
Article Title: Production scale-up and validation of packaging cell clearance of clinical-grade retroviral vector stocks produced in Cell Factories
Abstract: The clinical implementation of gene therapy requires large-scale production of viral vector stocks (VS) derived from packaging cell lines. Upon scaling-up, maintenance of high viral titers and filtration of the VS become significantly challenging. Thus, production schemes amenable to straightforward validation must be developed. To this end, we have established a semi-closed process to manufacture batches of 7l or more of clinical-grade oncoretroviral VS using 10-tray Cell Factories. Using a peristaltic pump, the VS are collected on 3 consecutive days, filtered, pooled and stored frozen. To ensure the absence of viable vector-producing cells (VPCs) from each VS unit-dose, we undertook an orthogonal log-removal validation study to demonstrate the ability of both the filtration system to remove viable cells and the VS freezing process to inactivate them. We demonstrate a total VPC-reduction of 11.6log, thus insuring the absence of contaminating VPCs in transduced clinical samples. We also show that this production process generates stable VS that can be stored at 80°C for more than 3 years. Importantly, this relatively simple and affordable process can be customized to generating large volume of VS for small animal or non-human primate studies. This methodology is not limited to the generation of cell-free clinical oncoretroviral VS, and can be applied to other types of vectors produced in packaging cell lines, such as lentiviral vectors. © 2006 Nature Publishing Group All rights reserved.
Keywords: nonhuman; validation process; methodology; quality control; animals; cell line; biotechnology; viral gene delivery system; animalia; genetic vectors; gene therapy; lentivirus; lentivirus vector; retrovirus vector; plasmids; cell separation; validation; bioreactors; scale-up; productivity; retroviridae; contamination; industry; virus titration; primates; freezing; viral gene therapy; clinical-grade retrovial vector; vector safety
Journal Title: Gene Therapy
Volume: 13
Issue: 1
ISSN: 0969-7128
Publisher: Nature Publishing Group  
Date Published: 2006-01-01
Start Page: 95
End Page: 100
Language: English
DOI: 10.1038/sj.gt.3302648
PUBMED: 16177816
PROVIDER: scopus
DOI/URL:
Notes: --- - "Cited By (since 1996): 10" - "Export Date: 4 June 2012" - "CODEN: GETHE" - "Source: Scopus"
Altmetric Score
MSK Authors
  1. Michel W J Sadelain
    462 Sadelain
  2. Isabelle C Riviere
    168 Riviere
  3. Joanna Hodges
    3 Hodges