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

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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 - Jan 1 2000

Fingerprint

Ovomucin
Carbohydrates
Light
Antigens
Antibodies
Glycoside Hydrolases
Complementarity Determining Regions
Glycosylation
DNA sequences
Consensus Sequence
Ovalbumin
Hybridomas
DNA Sequence Analysis
Lectins
Assays
Genes
Western Blotting
Monoclonal Antibodies
Cells

All Science Journal Classification (ASJC) codes

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

Cite this

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AU - Yamada, Koji

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