Effect of Culture Conditions on Glycosylation in Hybridoma Antibody Production

Abstract

To investigate the effect of culture conditions on glycosylation patterns, we carried out the batch cultivation (using serum-containing or serum-free media) and glucose-limited continuous cultivation (serum-free medium). Con A lectin blotting showed that N-linked glycosylation occurred under all the culture conditions used. The sugar composition of the oligosaccharide chain in each culture was studied using gas-chromatography and was found to be different under each culture condition. The detailed structure of each oligosaccharide chain was then analyzed by two-dimensional mapping techniques using pyridylamination method. The oligosaccharide chain content in human IgG was higher in batch cultivation using a serum medium than that in serum-free cultures. However, oligosaccharide chain content with a galactose residue at the chain end, which plays an important role in the immune system, was shown to be high in glucose-limited continuous cultivation under a low specific growth rate.