Process-scale purification from cell culture supernatants: monoclonal antibodies.
A method for processing monoclonal antibodies (mAb) from large volumes of cell culture supernatants using recently developed high performance and fast flow chromatography media is described. A high-antibody producing mouse hybridoma cell line was adapted to low serum containing medium (1% foetal calf serum) for the production of anti-tissue Plasminogen Activator (anti-tPA) monoclonal antibody (murine subclass IgG1). The process consisted of three main chromatographic steps: desalting, cation exchange on S Sepharose Fast Flow and gel filtration on Superose 6 prep grade. With this process for the purification of anti-tPA monoclonal antibody, the final product was greater than 95% pure with a total recovery of 75% i.e. 1.4 g was recovered in 0.345 l from 35 l of culture supernatant originally containing 1.9 g of mAb. The adaptation of this process for purification of other monoclonal antibodies is discussed.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D001709:Biotechnology / Q000379:methods; D002478:Cells, Cultured; D002845:Chromatography / Q000379:methods; D003470:Culture Media / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D006825:Hybridomas / Q000032:analysis*; D007074:Immunoglobulin G / Q000032:analysis; D051379:Mice; D010959:Tissue Plasminogen Activator / Q000276:immunology
D000911:Antibodies, Monoclonal; D003470:Culture Media; D007074:Immunoglobulin G; D010959:Tissue Plasminogen Activator