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Improved Fermentation Processes for NS0 Cell Lines Expressing Human Antibodies and Glutamine Synthetase
Author(s) -
Dempsey Jonathan,
Ruddock Steve,
Osborne Matthew,
Ridley Alison,
Sturt Simone,
Field Ray
Publication year - 2003
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp0256061
Subject(s) - transferrin , glutamine , antibody , cell culture , biochemistry , bovine serum albumin , amino acid , titer , biology , fed batch culture , glutamine synthetase , chemically defined medium , cell growth , microbiology and biotechnology , chemistry , fermentation , immunology , in vitro , genetics
Abstract To meet the increasing requirement for therapeutic antibodies to conduct clinical trials, an enhanced culture medium and fed‐batch process was developed for GS‐NS0 cell lines. This process was shown to produce high concentrations of monoclonal antibodies for several cell lines expressing different antibodies. Cells were adapted to growth in a glutamine‐ and serum‐free medium containing bovine serum albumin (BSA), cholesterol, and transferrin. A number of amino acids were found to be depleted during cell culture. The concentrations of these amino acids were increased, and further cell culture analyses were performed. This process of cell growth and analysis was repeated over multiple cycles until no depletion was detected. This resulted in an amino acid supplement that was shown to be generic and enhanced antibody productivity up to 5‐fold for the three cell lines tested. Transferrin was replaced using tropolone, a lipophilic iron chelator and ferric ammonium citrate. Cell growth was equivalent to that in transferrin‐containing medium over the wide ranges tested. A concentrated feed solution, based on the amino acid supplement and the components of the serum‐and protein‐free supplements, was formulated. Addition of this feed in response to metabolic requirements resulted in a harvest titer a further 2‐fold higher than the enhanced culture medium. Harvest antibody titers of up to 600 mg/L were achieved for three cell lines expressing different antibodies, representing an increase of 10‐fold over the starting concentrations.