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 1, 1991·Bio/technology·M J Page, M A Sydenham
Feb 1, 1992·Bio/technology·C R BebbingtonG T Yarranton
Jan 1, 1992·Cytotechnology·M E BrownT Hassell
Jan 24, 1991·Nature·G Winter, C Milstein
Mar 1, 1990·Tissue Antigens·G HaleH Waldmann
Dec 17, 1988·Lancet·G HaleH Waldmann
Mar 24, 1988·Nature·L RiechmannG Winter
Apr 1, 1988·Transplantation·G HaleH Waldmann
Sep 1, 1987·Cancer Genetics and Cytogenetics·E NachevaH Gadner
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
Dec 1, 1995·Glycobiology·M R LifelyJ Phillips
Oct 1, 1995·Cytotechnology·M J Keen, N T Rapson
Oct 1, 1995·Cytotechnology·M J Keen
Oct 1, 1995·Cytotechnology·M J Keen, T W Steward
Citations
Nov 13, 2008·Cytotechnology·Jinyou Zhang, David Robinson
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
Nov 13, 2008·Cytotechnology·L M BarnesA J Dickson
May 21, 2005·Biotechnology Progress·Erika Spens, Lena Häggström
Nov 15, 2011·Biotechnology Advances·Angela M MoraesCarlos A Pereira
May 22, 2010·Journal of Virological Methods·J Hobson-PetersP Toye
Apr 15, 2005·Biotechnology and Bioengineering·Gargi SethWei-Shou Hu
Apr 22, 2011·Biotechnology Progress·Yun WuSusan A Casnocha
Dec 31, 2005·Journal of Bioscience and Bioengineering·Shinobu KuwaeKaoru Kobayashi
Jan 4, 2013·PloS One·Sungwon JungBodour Salhia