title: A humanized anti-tumor necrosis factor-alpha monoclonal antibody that acts as a partial, competitive antagonist of the template antibody.
abstract: We have constructed several humanized versions of a monoclonal antibody (MAb78) against human tumor necrosis factor-alpha (huTNF-alpha) retaining the complementarity-determining regions (CDR) of the original mouse MAb with or without a variable number of original framework region (FR) residues. All versions, except one, showed a loss of binding affinity and neutralizing potency of at least 10-fold compared to the original mouse MAb or its chimeric equivalent. In some cases, however, the decrease in neutralizing potency was significantly greater than the decrease in binding affinity. Two humanized versions showing the greatest dissociation between these two parameters were studied for their capacity to inhibit the neutralizing activity of chimeric or murine MAb78 when used at concentrations that bound but only partially neutralized huTNF-alpha. One humanized version (MAb78D) was indeed able to do so, whereas the other (MAb78C) was not found to exert any inhibitory activity at all concentrations tested. The antagonistic effect of MAb78D was concentration dependent and could be overcome by increasing the concentrations of chimeric or murine MAb78. Two different models of MAb78-huTNF-alpha interaction that may help explain the antagonist activity of humanized MAb78D are discussed.
authors.affiliation: Scotgen Biopharmaceuticals Inc., Aberdeen, Scotland, UK.
authors.forename: P R
authors.initials: PR
authors.lastname: Tempest
authors.forename: E
authors.initials: E
authors.lastname: Barbanti
authors.forename: P
authors.initials: P
authors.lastname: Bremner
authors.forename: F J
authors.initials: FJ
authors.lastname: Carr
authors.forename: M
authors.initials: M
authors.lastname: Ghislieri
authors.forename: B
authors.initials: B
authors.lastname: Rifaldi
authors.forename: F
authors.initials: F
authors.lastname: Marcucci
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology*; D000915:Antibody Affinity; D001483:Base Sequence; D001667:Binding, Competitive / Q000276:immunology; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000276:immunology; D051379:Mice; D008969:Molecular Sequence Data; D009500:Neutralization Tests / Q000379:methods; D011993:Recombinant Fusion Proteins / Q000276:immunology; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D007135:Immunoglobulin Variable Region; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha