Procedure for obtaining recombinant mammalian myeloma cell lines adapted to growth in serum-free and protein-free media

摘要

Method for producing humanized monoclonal antibody hR3 directed against the EGF receptor or fragments thereof in serum-free media or proteins comprising adapting a recombinant mammalian myeloma cell line transfected with a sequence encoding said humanized monoclonal antibody hR3 directed against the receptor. EGF or fragments thereof, to a growth in medium without serum or proteins by a procedure comprising: I. A first stage in which the viability of the cell line is between 80 and 100% and the cells develop in media of culture with a consecutive reduction of the protein concentration until reaching a critical protein concentration at which cell viability decreases to 0%; and II. a second stage in which, once the critical concentration has been predetermined, the precritical concentration is set as a protein concentration at which cell growth is possible and is the starting point to slowly reduce the protein concentration to that the cell culture reaches the cell viability and the doubling time of the initial population.