The use of serum-free medium for the production of functionally active humanised monoclonal antibody from NS0 mouse myeloma cells engineered using glutamine synthetase as a selectable marker.

Cytotechnology
M J Keen, C Hale

Abstract

A protein-free growth medium (W38 medium) had previously been developed for the NS0 mouse myeloma cell line which is cholesterol-auxotrophic. This paper describes the development of a protein-free growth medium for NS0 cells expressing humanised monoclonal antibody using GS (glutamine synthetase) as a selectable marker. Several GS-engineered NS0 cell lines expressing humanised monoclonal antibody grew in a modification of W38 medium which maintained GS-selection, supplemented with cholesterol, phosphatidylcholine and β-cyclodextrin. Further studies showed that additional glutamic acid, asparagine, ribonucleosides and choline chloride improved cell growth. Amino acid analysis identified a number of amino acids that were being depleted from the culture medium. NS0 cell lines 9D4 and 2H5 expressing CAMPATH-1H(*) were adapted to enable them to grow serum-free in the absence of cholesterol and β-cyclodextrin. Cholesterol-independent 9D4 (9D4.CF) cells grown in shake flask culture using an enriched protein-free medium (WNSD medium), supplemented with human recombinant insulin (Nucellin), reached a maximum cell density to 1.86×10(6) cells ml(-1) producing 76.6 mg l(-1) of antibody. CAMPATH-1H antibody produced using serum-free medium ...Continue Reading

References

Sep 26, 1992·Lancet·J D IsaacsH Waldmann
Jan 24, 1991·Nature·G Winter, C Milstein
Mar 1, 1990·Tissue Antigens·G HaleH Waldmann
Mar 24, 1988·Nature·L RiechmannG Winter
Jun 1, 1987·The Journal of Experimental Medicine·J D SatoT Okamoto
Jul 1, 1980·Proceedings of the National Academy of Sciences of the United States of America·G Urlaub, L A Chasin

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Citations

Aug 13, 2008·Applied Biochemistry and Biotechnology·Suthasinee NilsangSudip Kumar Rakshit
Mar 6, 1998·Journal of Biotechnology·M DoverskogL Häggström
Apr 1, 1997·Current Opinion in Biotechnology·W S Hu, J G Aunins
Apr 15, 2005·Biotechnology and Bioengineering·Gargi SethWei-Shou Hu

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