High level expression of the humanized monoclonal antibody Campath-1H in Chinese hamster ovary cells.
We have cloned the light and heavy chain cDNAs for the humanized monoclonal antibody Campath-1H and expressed them in Chinese hamster ovary (CHO) cells using a dihydrofolate reductase (dhfr) amplification procedure. Each cDNA was positioned under control of the strong human beta-actin promoter/polyadenylation signals and used to evaluate alternative vector design and amplification procedures. By employing a dual selection co-transfection strategy, initial transformants accumulated antibody levels of 0.5 micrograms/ml after 4 days continuous culture. When subjected to successive rounds of selection in medium containing stepwise increments in methotrexate (MTX), stable cell lines were obtained that secreted up to 200 micrograms/ml of Campath-1H during the same period. This reflects a productivity of 100 micrograms/10(6) cells/day and demonstrates the potential of engineering CHO cells for the production of recombinant antibodies.
Department of Molecular Biology, Wellcome Biotech, Beckenham, Kent, UK.
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D000199:Actins / Q000235:genetics; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D000921:Antibody-Producing Cells / Q000378:metabolism; D015703:Antigens, CD / Q000276:immunology*; D000951:Antigens, Neoplasm; D000074301:CD52 Antigen; D002478:Cells, Cultured; D003001:Cloning, Molecular; D006224:Cricetinae; D003412:Cricetulus; D004247:DNA / Q000235:genetics; D005260:Female; D005784:Gene Amplification; D015870:Gene Expression; D005822:Genetic Vectors / Q000502:physiology; D006023:Glycoproteins; D006801:Humans; D007700:Kinetics; D009101:Multiple Myeloma / Q000235:genetics / Q000473:pathology; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D014407:Tumor Cells, Cultured
D000199:Actins; D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000951:Antigens, Neoplasm; D000074301:CD52 Antigen; C000613862:CD52 protein, human; C000613864:Cd52 protein, rat; D006023:Glycoproteins; D011994:Recombinant Proteins; D004247:DNA