Building high affinity human antibodies by altering the glycosylation on the light chain variable region in N-acetylglucosamine-supplemented hybridoma cultures.

Cytotechnology
H TachibanaS Shirahata

Abstract

We attempted to improve antibody affinity by varying glycosylation on the light chain variable region. The human hybridoma line HB4C5 produces an antibody reactive to lung adenocarcinoma, which possess a N-glycosylated carbohydrate chain on the light chain hypervariable region. It has been shown that altering this carbohydrate structure can be accomplished by varying the level of N-acetylglucosamine in glucose free medium, a change in the carbohydrate chain could be induced which resulted in modifying antigen binding. By culturing the cells in media containing more than 20 mM N-acetylglucosamine, cells produced antibody with 10 fold improved affinity as compared with antibody produced in 20 mM glucose-containing medium. A newly induced light chain glycoform produced in the N-acetylglucosamine-containing medium was shown to be responsible for this antigen binding enhancement. Addition of glucose in the N-acetylglucosamine-containing media led to decreased antibody affinity and slightly inhibited production of a new light chain in a dose-dependent manner. Combination of 20 mM N-acetylglucosamine and 0.5 mM glucose gave a higher antibody production without the decrease of the antigen binding. These results indicate that optimizati...Continue Reading

References

Apr 25, 1977·Biochemical and Biophysical Research Communications·N KoideT Muramatsu
Sep 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·H TowbinJ Gordon
Feb 1, 1979·Archives of Biochemistry and Biophysics·N J Stark, E C Heath
Dec 15, 1992·Biochemical and Biophysical Research Communications·H TachibanaH Murakami
Oct 1, 1988·Journal of Surgical Oncology·T YanoK Nomoto
Aug 20, 1987·Journal of Molecular Biology·C Chothia, A M Lesk
Oct 1, 1985·In Vitro Cellular & Developmental Biology : Journal of the Tissue Culture Association·H MurakamiH Omura
Jul 1, 1970·Proceedings of the National Academy of Sciences of the United States of America·H C Sox, L Hood
Feb 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·H MurakamiT Kano-Sueoka
Apr 4, 1964·Biochimica Et Biophysica Acta·J R CLAMPF W PUTNAM

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Citations

Jul 17, 2015·The Journal of Rheumatology·Walter P MaksymowychAnthony Marotta
Mar 10, 2019·The Journal of Immunology : Official Journal of the American Association of Immunologists·Jana KoersTheo Rispens
Jul 30, 2015·Biotechnology and Bioengineering·Helene Faustrup KildegaardMikael R Andersen
Feb 7, 2016·The Journal of Immunology : Official Journal of the American Association of Immunologists·Fleur S van de BovenkampYoann Rombouts
Feb 13, 2018·Proceedings of the National Academy of Sciences of the United States of America·Fleur S van de BovenkampTheo Rispens

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