Abstract
An anti-human tumor necrosis factor-alpha (TNF-alpha) monoclonal antibody, designated as 3B10, inhibits the biological activity of human TNF-alpha. In the present study, we constructed humanized version of the antibody by grafting its complementarity-determining regions (CDRs) onto a human antibody, HBS-1. Using a molecular model of mouse 3B10, framework residues affecting the CDR conformation were identified. Thus, these residues were also introduced into the framework together with the CDRs in a stepwise manner, depending on the degree of the possible importance of the residues. As a result, one humanized version (h3B10-9) which possesses nine mouse framework residues showed the same binding activity as that of the chimeric version. This humanized anti-TNF-alpha antibody is expected to be less immunogenic and thus more suitable for possible clinical use.
MeSH terms
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Amino Acid Sequence
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Animals
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Antibodies, Monoclonal / chemistry*
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Antibodies, Monoclonal / immunology*
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COS Cells / metabolism
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Cloning, Molecular
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DNA, Complementary / genetics
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DNA, Complementary / metabolism
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Enzyme-Linked Immunosorbent Assay
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Humans
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Immunoglobulin Variable Region / chemistry
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Immunoglobulin Variable Region / immunology
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Immunoglobulin Variable Region / metabolism
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Mice
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Molecular Sequence Data
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Neutralization Tests
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Polymerase Chain Reaction
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Protein Conformation
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Recombinant Fusion Proteins / chemical synthesis*
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Recombinant Fusion Proteins / immunology*
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Recombinant Fusion Proteins / metabolism
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Sequence Homology, Amino Acid
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Transfection
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Tumor Necrosis Factor-alpha / immunology*
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Tumor Necrosis Factor-alpha / metabolism
Substances
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Antibodies, Monoclonal
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DNA, Complementary
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Immunoglobulin Variable Region
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Recombinant Fusion Proteins
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Tumor Necrosis Factor-alpha