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
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Animals
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Antibodies, Monoclonal / chemistry*
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Antibodies, Monoclonal / genetics
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Antibodies, Monoclonal / immunology*
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Antibody-Dependent Cell Cytotoxicity*
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Antigens, Neoplasm / immunology*
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CHO Cells
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Cricetinae
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Fucose / chemistry*
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Fucosyltransferases / genetics
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Glycoproteins / immunology*
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Humans
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Immunoglobulin Constant Regions / genetics
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Immunoglobulin Constant Regions / immunology
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Immunoglobulin G / immunology
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Immunoglobulin Variable Region / genetics
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Immunoglobulin Variable Region / immunology
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Mutation
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Oligosaccharides / chemistry*
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Oligosaccharides / metabolism
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Receptors, IgG / immunology
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / immunology
Substances
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Antibodies, Monoclonal
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Antigens, Neoplasm
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Glycoproteins
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Immunoglobulin Constant Regions
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Immunoglobulin G
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Immunoglobulin Variable Region
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Oligosaccharides
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Receptors, IgG
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Recombinant Fusion Proteins
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tumor-associated antigen 72
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Fucose
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Fucosyltransferases
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Glycoprotein 6-alpha-L-fucosyltransferase