Large-scale clinical-grade retroviral vector production in a fixed-bed bioreactor Journal Article
| Authors: | Wang, X.; Olszewska, M.; Qu, J.; Wasielewska, T.; Bartido, S.; Hermetet, G.; Sadelain, M.; Rivière, I. |
| Article Title: | Large-scale clinical-grade retroviral vector production in a fixed-bed bioreactor |
| Abstract: | The successful genetic engineering of patient T cells with g-retroviral vectors expressing chimeric antigen receptors or T-cell receptors for phase II clinical trials and beyond requires the largescale manufacture of high-titer vector stocks. The production of retroviral vectors from stable packaging cell lines using roller bottles or 10-to 40-layer cell factories is limited by a narrow harvest window, labor intensity, open-system operations, and the requirement for significant incubator space. To circumvent these shortcomings, we optimized the production of vector stocks in a disposable fixed-bed bioreactor using good manufacturing practice-grade packaging cell lines. High-titer vector stocks were harvested over 10 days, representing a much broader harvest window than the 3-day harvest afforded by cell factories. For PG13 and 293Vec packaging cells, the average vector titer and the vector stocks' yield in the bioreactor were higher by 3.2-to 7.3-fold, and 5.6-to 13.1-fold, respectively, than those obtained in cell factories. The vector production was 10.4 and 18.6 times more efficient than in cell factories for PG13 and 293Vec cells, respectively. Furthermore, the vectors produced from the fixed-bed bioreactors passed the release test assays for clinical applications. Therefore, a single vector lot derived from 293Vec is suitable to transduce up to 500 patients cell doses in the context of large clinical trials using chimeric antigen receptors or T-cell receptors. These findings demonstrate for the first time that a robust fixed-bed bioreactor process can be used to produce g-retroviral vector stocks scalable up to the commercialization phase. © 2015 Wolters Kluwer Health, Inc. All rights reserved. |
| Keywords: | cell growth; cell line; retrovirus vector; cell density; phase 1 clinical trial (topic); scale up; process optimization; large scale production; good manufacturing practice; cell engineering; packaging; priority journal; article; fixed-bed bioreactor; g-retroviral vector; high vector titers; high vector yields; scalable clinical-grade vector manufacture; fixed bed reactor |
| Journal Title: | Journal of Immunotherapy |
| Volume: | 38 |
| Issue: | 3 |
| ISSN: | 1524-9557 |
| Publisher: | Lippincott Williams & Wilkins |
| Date Published: | 2015-04-01 |
| Start Page: | 127 |
| End Page: | 135 |
| Language: | English |
| DOI: | 10.1097/cji.0000000000000072 |
| PROVIDER: | scopus |
| PMCID: | PMC4353472 |
| PUBMED: | 25751502 |
| DOI/URL: | |
| Notes: | Export Date: 4 May 2015 -- Source: Scopus |
Altmetric Score
Related MSK Work