Changes of monosaccharide availability of human hybridoma lead to alteration of biological properties of human monoclonal antibody.
The effect of glucose and other monosaccharide availability in culture medium on production of antibody by human hybridomas has been studied. Human hybridoma cells C5TN produce an anti lung cancer human monoclonal antibody, and the light chain is N-glycosylated at the variable region. When the cell line was grown in the presence of various concentrations of glucose, the antibodies produced changed their antigen-binding activities. Analysis of the light chains produced under these condition revealed that four molecular-mass variant light chains ranging from about 26 to 32 kDa were secreted. The twenty six-kDa species, which corresponds to a non-glycosylated form of the light chain, was recovered after enzymatic removal of all N-linked carbohydrate chains, indicating that the source of the heterogeneity of the light chain is due to the varied glycosylation. When the C5TN cells were cultured in medium containing either fructose, mannose or galactose instead of glucose, galactose elevated the antigen binding activity of the antibody more than the other sugars. These results suggest that change of glucose availability affects the antigen-binding activity of the antibody via the alteration of the glycosylation.
Tachibana
H
H
Graduate School of Genetic Resources Technology, Kyushu University, Fukuoka, Japan.
Taniguchi
K
K
Ushio
Y
Y
Teruya
K
K
Osada
K
K
Murakami
H
H
D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000378:metabolism; D000937:Antigen-Antibody Reactions; D000941:Antigens / Q000378:metabolism; D002268:Carboxypeptidases / Q000276:immunology / Q000378:metabolism; D043422:Carboxypeptidases A; D003470:Culture Media; D003574:Cytochrome c Group / Q000276:immunology / Q000378:metabolism; D004247:DNA / Q000276:immunology / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D006825:Hybridomas / Q000378:metabolism*; D007147:Immunoglobulin Light Chains / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D008970:Molecular Weight; D009005:Monosaccharides / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D000941:Antigens; D003470:Culture Media; D003574:Cytochrome c Group; D007147:Immunoglobulin Light Chains; D009005:Monosaccharides; D004247:DNA; D002268:Carboxypeptidases; D043422:Carboxypeptidases A