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Production and characterization of human monoclonal antibody Fab fragments to vaccinia virus from a phage-display combinatorial library.


Immunoglobulin Fab Fragments, Animals, Bacteriophages, Leukocytes, Mononuclear, Humans, Mice, Amino Acid Sequence, Precipitin Tests, B-Lymphocytes, Vaccination, Cross Reactions, Antibodies, Monoclonal, Base Sequence, Vaccinia virus, immunology, Cercopithecus aethiops, Neutralization Tests, Molecular Sequence Data, Vero Cells, biosynthesis, Monkeypox virus, Gene Library

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      A combinatorial, phage-display library of human Fab antibody fragments was generated from IgG heavy chain (HC) and light chain (LC) genes cloned from the lymphocytes of a vaccinia virus (VACV)-immune donor. To ascertain the complexity of the library, nucleotide sequences of the variable regions of the HC and LC genes were determined. Fourteen distinct HC and 18 distinct LC (7 kappa and 11 lambda) that formed a combinatorial library of 22 Fabs were identified. Immune-precipitation of radiolabeled VACV revealed that at least six different VACV proteins were recognized by the antibodies. Plaque-reduction neutralization demonstrated that six of the Fabs neutralized VACV in the presence of anti-human antibody. ELISA studies indicated that 15 of the Fabs were cross-reactive with monkeypox virus.

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