Expression of GnTIII in a recombinant anti-CD20 CHO production cell line: Expression of antibodies with altered glycoforms leads to an increase in ADCC through higher affinity for FC gamma RIII

Biotechnol Bioeng. 2001 Aug 20;74(4):288-94.


The gene encoding the rat glycosylation enzyme beta1-4-N-acetylglucosaminyltransferase III (GnTIII) was cloned and coexpressed in a recombinant production Chinese hamster ovary (CHO) cell line expressing a chimeric mouse/human anti-CD20 IgG1 antibody. The new cell lines expressed high levels of antibody and have growth kinetics similar to that of the parent. Relative QPCR showed the cell lines to express varying levels of mRNA. High-performance liquid chromatography (HPLC) analysis showed the enzyme to have added bisecting N-acetylglucosamine (GlcNAc) residues in most (48% to 71%) of the N-linked oligosaccharides isolated from antibody preparations purified from the cell lines. In an ADCC assay the new antibody preparations promoted killing of CD20-positive target cells at approximately 10- to 20-fold lower concentrations than the parent. This activity was blocked using an anti-Fc gamma RIII antibody, supporting the role of Fc gamma RIII binding in this increase. In addition, cell binding assays showed the modified antibody bound better to Fc gamma RIII-expressing cells. The increase in ADCC activity is therefore likely due to an increased affinity of the modified antibody for the Fc gamma RIII receptor.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology
  • Antibody-Dependent Cell Cytotoxicity / immunology*
  • Antigens, CD20 / immunology*
  • CHO Cells
  • Carbohydrate Sequence
  • Cell Division
  • Chromatography, High Pressure Liquid
  • Cloning, Molecular
  • Cricetinae
  • Enzyme-Linked Immunosorbent Assay
  • Genetic Engineering
  • Glycosylation
  • Immunoglobulin G / chemistry*
  • Immunoglobulin G / genetics
  • Immunoglobulin G / immunology*
  • Immunoglobulin G / metabolism
  • Molecular Sequence Data
  • N-Acetylglucosaminyltransferases / genetics
  • N-Acetylglucosaminyltransferases / metabolism*
  • RNA, Messenger / analysis
  • RNA, Messenger / genetics
  • Rats
  • Receptors, IgG / immunology*
  • Receptors, IgG / metabolism
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology


  • Antibodies, Monoclonal
  • Antigens, CD20
  • Immunoglobulin G
  • RNA, Messenger
  • Receptors, IgG
  • Recombinant Proteins
  • N-Acetylglucosaminyltransferases
  • beta-1,4-mannosyl-glycoprotein beta-1,4-N-acetylglucosaminyltransferase