Fucose removal from complex-type oligosaccharide enhances the antibody-dependent cellular cytotoxicity of single-gene-encoded antibody comprising a single-chain antibody linked the antibody constant region

J Immunol Methods. 2005 Nov 30;306(1-2):93-103. doi: 10.1016/j.jim.2005.07.025. Epub 2005 Oct 3.

Abstract

Fucose removal from complex-type oligosaccharide of human IgG1-type antibody results in a great enhancement of antibody-dependent cellular cytotoxicity (ADCC). The aim of this study was to clarify the effect of fucose removal on effector functions of a single-gene-encoded antibody with an scFv used as the binding domain. We generated both a fucose-negative anti-tumor associated glycoprotein (TAG)-72 scFv-Fc using alpha-1,6-fucosyltransferase knock-out CHO cells and a highly fucosylated scFv-Fc from parental CHO cells. Expression, assembly and antigen binding activity of the scFv-Fcs were not influenced by fucose removal. The scFv-Fc lacking fucose exhibited significantly more potent FcgammaRIIIa binding and ADCC compared to highly fucosylated scFv-Fc. These results prove that ADCC enhancement by fucose-removal is effective in not only whole IgG1, but also scFv-Fc, and thus increases the potential of Fc-fusion proteins as therapeutic candidates.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / chemistry*
  • Antibodies, Monoclonal / genetics
  • Antibodies, Monoclonal / immunology*
  • Antibody-Dependent Cell Cytotoxicity*
  • Antigens, Neoplasm / immunology*
  • CHO Cells
  • Cricetinae
  • Fucose / chemistry*
  • Fucosyltransferases / genetics
  • Glycoproteins / immunology*
  • Humans
  • Immunoglobulin Constant Regions / genetics
  • Immunoglobulin Constant Regions / immunology
  • Immunoglobulin G / immunology
  • Immunoglobulin Variable Region / genetics
  • Immunoglobulin Variable Region / immunology
  • Mutation
  • Oligosaccharides / chemistry*
  • Oligosaccharides / metabolism
  • Receptors, IgG / immunology
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / immunology

Substances

  • Antibodies, Monoclonal
  • Antigens, Neoplasm
  • Glycoproteins
  • Immunoglobulin Constant Regions
  • Immunoglobulin G
  • Immunoglobulin Variable Region
  • Oligosaccharides
  • Receptors, IgG
  • Recombinant Fusion Proteins
  • tumor-associated antigen 72
  • Fucose
  • Fucosyltransferases
  • Glycoprotein 6-alpha-L-fucosyltransferase