Optimization of a transient antibody expression platform towards high titer and efficiency.
CHO cells
antibody
drug discovery
titer
transient transfection
Journal
Biotechnology journal
ISSN: 1860-7314
Titre abrégé: Biotechnol J
Pays: Germany
ID NLM: 101265833
Informations de publication
Date de publication:
Apr 2021
Apr 2021
Historique:
revised:
10
11
2020
received:
29
05
2020
pubmed:
24
11
2020
medline:
28
4
2021
entrez:
23
11
2020
Statut:
ppublish
Résumé
Transient gene expression (TGE) using mammalian cells is an extensively used technology for the production of antibodies and recombinant proteins and has been widely adopted by both academic and industrial labs. Chinese Hamster Ovary (CHO) cells have become one of the major workhorses for TGE of recombinant antibodies due to their attractive features: post-translational modifications, adaptation to high cell densities, and use of serum-free media. In this study, we describe the optimization of parameters for TGE for antibodies from CHO cells. Through a matrix evaluation of multiple factors including inoculum, transfection conditions, amount and type of DNA used, and post-transfection culture conditions, we arrived at an uniquely optimized process with higher titer and reduced costs and time, thus increasing the overall efficiency of early antibody material supply. We further investigated the amount of coding DNA used in TGE and the influence of kinetics and size of the transfection complex on the in vitro efficiency of the transfection. We present here the first report of an optimized TGE platform using Filler DNA in an early drug discovery setting for the screening and production of therapeutic mAbs.
Identifiants
pubmed: 33226178
doi: 10.1002/biot.202000251
doi:
Substances chimiques
Recombinant Proteins
0
Polyethyleneimine
9002-98-6
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
e2000251Informations de copyright
© 2021 Wiley-VCH GmbH.
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