Production and purification of a bladder cancer associated membrane protein

ABSTRACT The aim of the study was to produce an antigen, mainly confined to bladder carcinomas, by use of cell culture in a pilot scale fermenter. Human bladder cancer cells, TCCSuP, grown on Cytodex R 1 were scaled up by serial subcultlvation to a final culture volume of 40 litres. The cell doubling time was reduced during the scale up process by replacing the NBS culture supplement with FBS. An increased cell growth rate was obtained in a specially designed pilot scale fermenter, as compared to growth rates obtained in spinner flasks, by carefully controlling the pH, dissolved oxygen and stirring speed. Following a eel1-microcarrier separation step the antigen was purified from a cell membrane preparation. The monoclonal antibody, 7E9, which was earlier produced against TCCSuP cells, was used to define the antigen under study. The antigen was further used to obtain polyclonal antisera from immunised rabbits. This antisera is now being used in developing a clinical assay for the detection of bladder carcinoma antigen present in the urine of affected persons.