N-linked glycan structures of a mouse monoclonal antibody produced from tobacco BY2 suspension-cultured cells

J Biosci Bioeng. 2006 Mar;101(3):212-8. doi: 10.1263/jbb.101.212.

Abstract

cDNA encoding H- and L-chains from a mouse monoclonal antibody was introduced into tobacco BY2 cells, and the resulting sugar chain structures of plant-produced antibodies were analyzed by a combination of HPLC, exoglucosidase digestion and matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. The glycan structures determined were Man(5-6)GlcNAc2 (22.3%), GlcNAcMan5GlcNAc2 (3.1%), GlcNAcMan3FucXylGlcNAc2 (24.4%), GlcNAcMan3XylGlcNAc2 (17.8%), Man3FucXylGlcNAc2 (24.3%), and Man3XylGlcNAc2 (8.1%). The major glycan structures of the antibodies produced by transgenic suspension-cultured cells contain typical plant bisecting beta(1,2)-xylose and alpha(1,3)-fucose residues, suggesting the posttranslational modification of a recombinant antibody in the late Golgi apparatus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / chemistry*
  • Base Sequence
  • Carbohydrate Sequence
  • Cloning, Molecular
  • Genes, Plant
  • Glucosidases / metabolism
  • Golgi Apparatus / metabolism
  • Immunoglobulin G / chemistry
  • Mice
  • Molecular Sequence Data
  • Nicotiana / metabolism*
  • Plants / chemistry
  • Plants / metabolism
  • Polysaccharides / chemistry*

Substances

  • Antibodies, Monoclonal
  • Immunoglobulin G
  • Polysaccharides
  • Glucosidases