Rat monoclonal antibodies. IV. Easy method for in vitro production

A simple technique for in vitro production of monoclonal antibodies is described. Two to 5 X 10(6) hybridoma cells are diluted in 300 ml of medium in a 2 litre 'roller bottle' and incubated for 2 weeks without subsequent adjustment of the cell concentration or renewal of the medium. The method is based on the observation that when hybridoma cell cultures are maintained in the same medium, although cell growth stops after a few days and the percentage of dead cells increases rapidly, the amount of monoclonal antibody present in the culture medium increases up to the 15th day, the yield reaching more than 10 mg per flask. The method can be used both for rat and mouse hybridoma cell culture.