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.