Lack of fucose on human IgG1 N-linked oligosaccharide improves binding to human Fcgamma RIII and antibody-dependent cellular toxicity.
Lec13 cells, a variant Chinese hamster ovary cell line, were used to produce human IgG1 that were deficient in fucose attached to the Asn(297)-linked carbohydrate but were otherwise similar to that found in IgG1 produced in normal Chinese hamster ovary cell lines and from human serum. Lack of fucose on the IgG1 had no effect on binding to human FcgammaRI, C1q, or the neonatal Fc receptor. Although no change in affinity was found for the His(131) polymorphic form of human FcgammaRIIA, a slight improvement in binding was evident for FcgammaRIIB and the Arg(131) FcgammaRIIA polymorphic form. In contrast, binding of the fucose-deficient IgG1 to human FcgammaRIIIA was improved up to 50-fold. Antibody-dependent cellular cytotoxicity assays using purified peripheral blood monocytes or natural killer cells from several donors showed enhanced cytotoxicity, especially evident at lower antibody concentrations. When combined with an IgG1 Fc protein variant that exhibited enhanced antibody-dependent cellular cytotoxicity, the lack of fucose was synergistic.
Shields
Robert L
RL
Department of Immunology, Genentech, Inc., 1 South San Francisco, CA 94080, USA.
Lai
Jadine
J
Keck
Rodney
R
O'Connell
Lori Y
LY
Hong
Kyu
K
Meng
Y Gloria
YG
Weikert
Stefanie H A
SH
Presta
Leonard G
LG
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D001216:Asparagine / Q000378:metabolism; D016466:CHO Cells; D002241:Carbohydrates / Q000737:chemistry; D002460:Cell Line; D006224:Cricetinae; D018076:DNA, Complementary / Q000378:metabolism; D019281:Dimerization; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D005643:Fucose / Q000737:chemistry*; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D011110:Polymorphism, Genetic; D011485:Protein Binding; D017452:Receptors, IgG / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014162:Transfection
D000911:Antibodies, Monoclonal; D002241:Carbohydrates; D018076:DNA, Complementary; D007074:Immunoglobulin G; D017452:Receptors, IgG; D005643:Fucose; D001216:Asparagine