High-level expression of a recombinant antibody from myeloma cells using a glutamine synthetase gene as an amplifiable selectable marker.
作者： C. R. Bebbington , G. Renner , S. Thomson , D. King , D. Abrams
DOI: 10.1038/NBT0292-169
关键词:
摘要: We report a method for introducing glutamine synthetase (GS) selectable marker into myeloma cells in which transfectants are selected by growth glutamine-free medium. Vector amplification can subsequently be using the specific inhibitor of GS, methionine sulphoximine (MSX). Using this system, DNA sequences encoding chimeric B72.3 IgG4 antibody were expressed from hCMV-MIE promoters NSO cells. A cell line was isolated after single round selection vector contains approximately 4 copies vector, secretes 10-15 pg/cell/day cB72.3 during exponential and accumulate 560 mg/l fed-batch air-lift fermentation system. Productivity is stable absence MSX selection.
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sci-hub.se 参考文章(43)
S.L. Bell, C.R. Bebbington, M.E. Bushell, P.G. Sanders, M.F. Scott, R.E. Spier, J.N. Wardell, GENETIC ENGINEERING OF CELLULAR PHYSIOLOGY Production of Biologicals from Animal Cells in Culture. pp. 304- 308 ,(1991) , 10.1016/B978-0-7506-1103-9.50062-7
Rafael Palacios, Jaime Mora, Glutamine : metabolism, enzymology, and regulation Academic Press. ,(1980)
G. Galfrè, C. Milstein, [1] Preparation of monoclonal antibodies: Strategies and procedures Methods in Enzymology. ,vol. 73, pp. 3- 46 ,(1981) , 10.1016/0076-6879(81)73054-4
Harry Eagle, Vance I. Oyama, Mina Levy, Clara L. Horton, Ralph Fleischman, THE GROWTH RESPONSE OF MAMMALIAN CELLS IN TISSUE CULTURE TO l-GLUTAMINE AND l-GLUTAMIC ACID Journal of Biological Chemistry. ,vol. 218, pp. 607- 616 ,(1956) , 10.1016/S0021-9258(18)65826-0
D V Morrissey, B A Stone, C A Guindon, H Dorai, G L Davis, J Saltzgaber-Muller, F Toneguzzo, S P Lilly, K W McDonald, B G Sahagan, A genetically engineered murine/human chimeric antibody retains specificity for human tumor-associated antigen. Journal of Immunology. ,vol. 137, pp. 1066- 1074 ,(1986)
John E. Shively, Arthur D. Riggs, Michael Neumaier, Raymond J. Paxton, Franz-Josef Gaida, Louise Shively, Christine Ilgen, Fun-Shan Chen, Cloning of the genes for T84.66, an antibody that has a high specificity and affinity for carcinoembryonic antigen, and expression of chimeric human/mouse T84.66 genes in myeloma and Chinese hamster ovary cells. Cancer Research. ,vol. 50, pp. 2128- 2134 ,(1990)
C B Beidler, B W Unger, J Phelps, L J Myers, J R Ludwig, C G Papworth, J Cardenas, E Melcher, M Sierzega, M Fisher, Cloning and high level expression of a chimeric antibody with specificity for human carcinoembryonic antigen. Journal of Immunology. ,vol. 141, pp. 4053- 4060 ,(1988)
I Hellström, E D Murray, K E Hellström, J A Ledbetter, R R Robinson, A Y Liu, Production of a mouse-human chimeric monoclonal antibody to CD20 with potent Fc-dependent biologic activity. Journal of Immunology. ,vol. 139, pp. 3521- 3526 ,(1987)
C R Wood, D Wilson, R M O'Hara, A J Dorner, E M Alderman, R J Kaufman, G E Morris, High level synthesis of immunoglobulins in Chinese hamster ovary cells. Journal of Immunology. ,vol. 145, pp. 3011- 3016 ,(1990)
D J King, J R Adair, S Angal, D C Low, K A Proudfoot, J C Lloyd, M W Bodmer, G T Yarranton, Expression, purification and characterization of a mouse-human chimeric antibody and chimeric Fab' fragment. Biochemical Journal. ,vol. 281, pp. 317- 323 ,(1992) , 10.1042/BJ2810317