Rapid Lentiviral Vector Producer Cell Line Generation Using a Single DNA Construct.
cell line development
lentiviral vector
manufacturing
producer cell line
Journal
Molecular therapy. Methods & clinical development
ISSN: 2329-0501
Titre abrégé: Mol Ther Methods Clin Dev
Pays: United States
ID NLM: 101624857
Informations de publication
Date de publication:
11 Dec 2020
11 Dec 2020
Historique:
received:
28
04
2020
accepted:
07
08
2020
entrez:
30
9
2020
pubmed:
1
10
2020
medline:
1
10
2020
Statut:
epublish
Résumé
Stable suspension producer cell lines for the production of vesicular stomatitis virus envelope glycoprotein (VSVg)-pseudotyped lentiviral vectors represent an attractive alternative to current widely used production methods based on transient transfection of adherent 293T cells with multiple plasmids. We report here a method to rapidly generate such producer cell lines from 293T cells by stable transfection of a single DNA construct encoding all lentiviral vector components. The resulting suspension cell lines yield titers as high as can be achieved with transient transfection, can be readily scaled up in single-use stirred-tank bioreactors, and are genetically and functionally stable in extended cell culture. By removing the requirement for efficient transient transfection during upstream processing of lentiviral vectors and switching to an inherently scalable suspension cell culture format, we believe that this approach will result in significantly higher batch yields than are possible with current manufacturing processes and enable better patient access to medicines based on lentiviral vectors.
Identifiants
pubmed: 32995359
doi: 10.1016/j.omtm.2020.08.011
pii: S2329-0501(20)30177-7
pmc: PMC7501408
doi:
Types de publication
Journal Article
Langues
eng
Pagination
47-57Informations de copyright
© 2020 The Authors.
Références
Science. 1989 Jun 16;244(4910):1307-12
pubmed: 2660262
Nucleic Acids Res. 2013 Apr;41(8):e92
pubmed: 23444141
Hum Gene Ther. 2001 Mar 1;12(4):359-65
pubmed: 11242528
J Biotechnol. 2016 Dec 20;240:23-30
pubmed: 27769802
J Virol. 1998 Nov;72(11):8463-71
pubmed: 9765382
J Virol. 2011 Jul;85(13):6205-11
pubmed: 21507965
Mol Ther Methods Clin Dev. 2016 Apr 13;3:16017
pubmed: 27110581
EMBO Mol Med. 2017 Nov;9(11):1558-1573
pubmed: 28835507
Gene Ther. 2009 Jun;16(6):805-14
pubmed: 19262613
Blood. 2009 May 21;113(21):5104-10
pubmed: 19286997
Methods Mol Biol. 2016;1418:335-51
pubmed: 27008022
Mol Ther. 2002 Mar;5(3):242-51
pubmed: 11863413
Expert Opin Biol Ther. 2013 Jul;13(7):987-1011
pubmed: 23590247
Mol Ther Methods Clin Dev. 2016 May 11;3:16033
pubmed: 27222840
Biotechnol Adv. 2015 Dec;33(8):1878-96
pubmed: 26523782
Nat Biotechnol. 2003 May;21(5):569-72
pubmed: 12679787
Biotechnol J. 2013 Jul;8(7):811-21
pubmed: 23450727
Hum Gene Ther. 1998 Sep 1;9(13):1939-50
pubmed: 9741432
Virology. 2000 Jan 5;266(1):170-9
pubmed: 10612671
Hum Vaccin Immunother. 2013 Nov;9(11):2398-408
pubmed: 24056737
Hum Gene Ther. 2011 Mar;22(3):357-69
pubmed: 21070114
Cell. 1981 Jan;23(1):29-39
pubmed: 7214526
Science. 2013 Aug 23;341(6148):1233158
pubmed: 23845948
Cytotechnology. 2006 Mar;50(1-3):9-33
pubmed: 19003068
Genome Res. 2002 Sep;12(9):1434-44
pubmed: 12213781
Cytotechnology. 2013 Oct;65(5):791-3
pubmed: 23975256
Nat Commun. 2014 Sep 03;5:4767
pubmed: 25182477
PLoS One. 2008;3(11):e3647
pubmed: 18985154
J Mol Biol. 2007 Nov 30;374(3):580-90
pubmed: 17945255
Proc Natl Acad Sci U S A. 2008 Jul 29;105(30):10547-52
pubmed: 18650378
Science. 2001 Dec 14;294(5550):2368-71
pubmed: 11743206
J Virol. 1998 Apr;72(4):2663-70
pubmed: 9525583
Nat Methods. 2017 Apr;14(4):417-419
pubmed: 28263959
Nat Commun. 2017 Mar 27;8:14834
pubmed: 28345582
Nat Biotechnol. 2014 Oct;32(10):1019-25
pubmed: 25129690