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Generation of a cholesterol‐independent, non‐GS NS0 cell line through chemical treatment and application for high titer antibody production
Author(s) -
Li Jincai,
Gu Wei,
Edmondson Diane G.,
Lu Connie,
Vijayasankaran Natarajan,
Figueroa Bruno,
Stevenson Dave,
Ryll Thomas,
Li Feng
Publication year - 2012
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.24450
Subject(s) - clone (java method) , titer , auxotrophy , cholesterol , recombinant dna , cell culture , biology , microbiology and biotechnology , western blot , plasmid , biochemistry , transfection , host (biology) , chemistry , antibody , gene , genetics , escherichia coli
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. Biotechnol. Bioeng. 2012; 109:1685–1692. © 2012 Wiley Periodicals, Inc.