Format

Send to

Choose Destination
See comment in PubMed Commons below
Biotechnol Bioeng. 2012 Jul;109(7):1685-92. doi: 10.1002/bit.24450. Epub 2012 Feb 6.

Generation of a cholesterol-independent, non-GS NS0 cell line through chemical treatment and application for high titer antibody production.

Author information

1
Oceanside Pharma Technical Development, Genentech, Inc, Oceanside, California, USA.

Abstract

NS0 cells require exogenous cholesterol for growth. The non-glutamine synthetase (GS) cholesterol-dependent NS0 host was treated with 5-azacytidine (5azaC), a demethylation drug, and adapted to grow in cholesterol-free, chemically defined medium. Within 7 weeks, a stable, cholesterol-independent NS0 host (NS0.CF) was obtained. The new NS0.CF host, as well as the original cholesterol auxotroph host, was transfected with the same mAb expression plasmid, and the top producing clone from both hosts were compared side-by-side in the enhanced platform fed-batch cultures using chemically defined media. The NS0.CF derived clone significantly out-performed the cholesterol-dependent clone, with titer reaching 4.5 g/L versus 3.0 g/L, respectively, mainly due to higher specific productivity, while key product quality attributes remained comparable. This work demonstrated an effective and rapid approach to generate a cholesterol-independent NS0 host, and its application in recombinant protein production.

PMID:
22252532
DOI:
10.1002/bit.24450
[Indexed for MEDLINE]
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for Wiley
    Loading ...
    Support Center