title: High yield of human monoclonal antibody produced by stably transfected Drosophila schneider 2 cells in perfusion culture using wave bioreactor.
abstract: Since it was first introduced in late 1990s Wave bioreactor has been used for protein production by mammalian and insect cell lines. However, using Wave bioreactor to produce human monoclonal antibody by stable Drosophila Schneider 2 (S2) cell transfectants has not been reported before. In this study, S2 cells were co-transfected with an inducible vector expressing human monoclonal antibody heavy and light chains, respectively, specific for hemagglutinin (HA) of H5N1 influenza virus. Stable S2 transfectant clone was selected by limiting dilution assay. Stable S2 transfectant clone that produce the highest amount of human monoclonal antibody was inoculated into two 2-l disposable cellbags, where cell growth and antibody production were compared between batch and perfusion cultures using Wave bioreactor. Here, we report that maximum viable cell density reached 1.06 × 10(7) cells/ml in batch culture; whereas 1.04 × 10(8) cells/ml was achieved in perfusion culture. The maximum volumetric antibody productivity in batch culture was 52 mg/l/day; while perfusion culture yielded 1,437 mg/l/day. As a result, the total antibody production was 201 mg in batch culture and 8,212 mg in perfusion culture. The antibody produced by both cultures displays full neutralizing activity. Thus, our results provide strong support for using Wave bioreactor in perfusion culture for a large-scale production of human monoclonal antibody by stable S2 cell transfectants.
authors.affiliation: Unit of Anti-Viral Immunity and Genetic Therapy, Key Laboratory of Molecular Virology and Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai 200025, China.
authors.forename: Lulan
authors.initials: L
authors.lastname: Wang
authors.forename: Hongxing
authors.initials: H
authors.lastname: Hu
authors.forename: Jianjun
authors.initials: J
authors.lastname: Yang
authors.forename: Feng
authors.initials: F
authors.lastname: Wang
authors.forename: Christian
authors.initials: C
authors.lastname: Kaisermayer
authors.forename: Paul
authors.initials: P
authors.lastname: Zhou
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D000914:Antibodies, Viral / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D019149:Bioreactors; D001709:Biotechnology; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D004330:Drosophila / Q000145:classification / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000276:immunology; D006801:Humans; D053124:Influenza A Virus, H5N1 Subtype / Q000276:immunology*; D010477:Perfusion; D014162:Transfection
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D019267:Hemagglutinin Glycoproteins, Influenza Virus