Development of optimized transfectoma cell lines for production of chimeric antibodies in hollow fiber cell culture systems

✍ Scribed by Beatrice S. Schläpfer; Marcel Scheibler; Anke-Peggy Holtorf; Hai Van Nguyen; Gerd Pluschke

• Publisher: John Wiley and Sons
• Year: 1995
• Tongue: English
• Weight: 987 KB
• Volume: 45
• Category: Article
• ISSN: 0006-3592
• DOI: 10.1002/bit.260450405

✦ Synopsis

Methods for the selection of transfectoma cells that express large quantities of mouse-human chimeric antibodies have been developed. SP2/0 mouse myeloma cells were transfected with pSV2-gpt and pSV2-neo based immunoglobulin expression vectors. Double transfectants were selected using the xanthineguanine phosphoribosyl transferase (gpt) and the neomycin (neo) selection marker genes. ELISA-based screening of transfectoma clones resulted in the isolation of IgG-producing transfectomas. Introduction of the kappa light-chain 3'enhancer into the light-chain expression vector significantly increased immunoglobulin expression, but only when the enhancer was located at its physiological site, 9 kb downstream of the kappa constant region exon.

With some of the transfectomas, final yields of up to 80 mg/L of chimeric IgG were obtained in conventional flask cultures using serum-free growth medium. A pilot-scale AcuSyst Maximizer hollow fiber cell culture system was used for the production of gram amounts of chimeric IgG. Results obtained with different transfectoma clones in conventional culture were not fully predictive for yields in the hollow fiber system. In contrast, differences in productivity between individual clones in the laboratory-scale Tecnomouse cell culture unit were comparable with those in the Maximizer system. Up to 200 mg of chimeric IgG were produced per day in one Maximizer bioreactor.