Structural analysis of the carbohydrate chains of a mouse monoclonal IgM antibody

Eur J Biochem. 1989 Sep 1;184(1):29-38. doi: 10.1111/j.1432-1033.1989.tb14986.x.

Abstract

A mouse monoclonal IgM antibody, directed against human blood group B determinant, was isolated from hybridoma culture growth medium. Chemical analysis indicated presence of N- and O-linked oligosaccharides. The N- and O-linked carbohydrate chains were liberated using two different conditions of reductive alkaline degradation. Structural analysis was carried out on the isolated chains using chemical analysis, 500-MHz 1H-NMR spectroscopy and fast-atom-bombardment mass spectrometry. The following composite structures of the N-linked chains were found: (formula; see text) where R = OH for biantennary structures and R = Neu5Ac alpha 2-3Gal beta 1-4 GlcNAc beta 1- or Neu5Ac alpha 2-3Gal beta 1-3[Neu5Ac alpha 2-6]GlcNAc beta 1- for triantennary structures. The O-linked oligosaccharides, found in the light chains, were shown to have the structure Neu5Ac alpha 2-3Gal beta 1-3GalNAc. The native IgM antibody could be separated on a concanavalin-A-Sepharose column into two subfractions, differing in the presence of a high-mannose-type oligosaccharide.

MeSH terms

  • ABO Blood-Group System / immunology
  • Animals
  • Antibodies, Monoclonal* / isolation & purification
  • Carbohydrate Conformation
  • Carbohydrate Sequence
  • Humans
  • Immunoglobulin M* / isolation & purification
  • Magnetic Resonance Spectroscopy
  • Mass Spectrometry
  • Mice
  • Oligosaccharides / isolation & purification

Substances

  • ABO Blood-Group System
  • Antibodies, Monoclonal
  • Immunoglobulin M
  • Oligosaccharides