Antigen binding of an ovomucoid-specific antibody is affected by a carbohydrate chain located on the light chain variable region

Yoshinori Fujimura, Hirofumi Tachibana, Nozomu Eto, Koji Yamada

Research output: Contribution to journalArticlepeer-review

17 Citations (Scopus)

Abstract

We cloned the variable regions of heavy and light chain genes of an anti-ovomucoid monoclonal antibody (MAb-OM21) produced by the mouse hybridoma cell line OM21. DNA sequence analysis showed that the light chain of the MAb-OM21 has only one potential N-glycosylation consensus sequence in the complementarity determining region 2 of the light chain. To find whether carbohydrate chains are located on the light chain, we assayed for the size of the light chain, after treatment with N-glycosidase, by western blotting, and also detection of the carbohydrate chains on the light chain was done using the lectin blot assay. A N-linked carbohydrate chain has been shown to bind to the light chain. To clarify the role of this carbohydrate chain in the light chain, we produced carbohydrate variant antibodies by N-deglycosylation using glycosidase or by expressing the antibody from different host cells. The N-deglycosylated variant antibody has greater antigen binding, and the antibody produced from the different host cells showed a reduced antigen binding activity and acquired the ability to react to ovalbumin. These results suggest that antigen binding of the ovomucoid specific antibody MAb-OM21 can be affected by the carbohydrate chain on the light chain variable region.

Original languageEnglish
Pages (from-to)2298-2305
Number of pages8
JournalBioscience, Biotechnology and Biochemistry
Volume64
Issue number11
DOIs
Publication statusPublished - 2000

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

Fingerprint Dive into the research topics of 'Antigen binding of an ovomucoid-specific antibody is affected by a carbohydrate chain located on the light chain variable region'. Together they form a unique fingerprint.

Cite this