Abstract
A human recombinant monoclonal Fab fragment that specifically recognizes all the influenza A virus strains tested was produced in transformed Escherichia coli using the phage display technique. No strain of influenza B virus reacted with it. It was purified after four cycles of panning and by a single passage through an immunoaffinity column. About 1 mg of pure monoclonal antibody was obtained from 1 liter of culture medium in 3 working days. The Fab fragment reacted with a viral 27-kDa protein, which could reasonably be a matrix protein. Indirect immunofluorescence tests performed on virus-infected MDCK cells showed that this Fab fragment was at least equally efficient as other commercial monoclonal antibody-based systems in detecting influenza A viral infections. The potential advantages of human recombinant Fabs on murine monoclonal antibodies are discussed.
Publication types
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Research Support, Non-U.S. Gov't
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Validation Study
MeSH terms
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Alphainfluenzavirus / immunology*
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Amino Acid Sequence
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Animals
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Antibodies, Monoclonal / genetics
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Antibodies, Monoclonal / immunology*
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Antibody Specificity
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Antigens, Viral / immunology
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Cell Line
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Dogs
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Escherichia coli
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Fluorescent Antibody Technique, Indirect
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Humans
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Immunoglobulin Fab Fragments / genetics
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Immunoglobulin Fab Fragments / immunology*
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Immunoglobulin Heavy Chains / immunology
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Immunoglobulin kappa-Chains / immunology
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Immunoglobulin lambda-Chains / immunology
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Kidney
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Molecular Sequence Data
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / immunology
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Sensitivity and Specificity
Substances
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Antibodies, Monoclonal
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Antigens, Viral
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Immunoglobulin Fab Fragments
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Immunoglobulin Heavy Chains
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Immunoglobulin kappa-Chains
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Immunoglobulin lambda-Chains
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Recombinant Fusion Proteins