title: Recombinant human monoclonal antibody IgG1b12 neutralizes diverse human immunodeficiency virus type 1 primary isolates.
abstract: The CD4-binding domain of human immunodeficiency virus type 1 (HIV-1) gp120 elicits antibodies that are present in infected human sera. Monoclonal antibodies that recognize the HIV-1 gp120 CD4-binding domain have been isolated. Some of these antibodies can neutralize laboratory-adapted strains of HIV-1 and probably mediate neutralization by interfering with virus binding to its cellular CD4 receptor. However, most anti-CD4 binding domain antibodies do not neutralize primary HIV-1 isolates. We used primary HIV-1 isolates in an infectivity reduction assay to test the uniquely derived anti-CD4 binding domain recombinant human monoclonal antibody, IgG1b12. All of the tested HIV-1 isolates were neutralized by this antibody. Additional studies indicated that neutralization of a primary isolate with MAb IgG1b12 did not require continuous exposure of human peripheral blood mononuclear cell cultures to the antibody. Finally, a complete IgG1 molecule of an in vitro-selected b12 FAb mutant with a > 400-fold increase in affinity was assembled, expressed in mammalian cells, and evaluated in the infectivity reduction assay in comparative studies with the parent IgG1b12 antibody. The mutant did not retain the level of primary isolate neutralization potency that was a property of the parent molecule. Thus, we confirm that recombinant IgG1b12 has a unique specificity, and that it can neutralize all primary isolates tested in human PBMC cultures in vitro.
authors.affiliation: Department of Antiviral Research, Merck Research Laboratories, West Point, Pennsylvania 19486, USA.
authors.forename: J A
authors.initials: JA
authors.lastname: Kessler
authors.forename: P M
authors.initials: PM
authors.lastname: McKenna
authors.forename: E A
authors.initials: EA
authors.lastname: Emini
authors.forename: C P
authors.initials: CP
authors.lastname: Chan
authors.forename: M D
authors.initials: MD
authors.lastname: Patel
authors.forename: S K
authors.initials: SK
authors.lastname: Gupta
authors.forename: G E
authors.initials: GE
authors.lastname: Mark
authors.forename: C F
authors.initials: CF
authors.lastname: Barbas
authors.forename: D R
authors.initials: DR
authors.lastname: Burton
authors.forename: A J
authors.initials: AJ
authors.lastname: Conley
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology*; D015704:CD4 Antigens / Q000276:immunology; D002478:Cells, Cultured; D015699:HIV Envelope Protein gp120 / Q000235:genetics* / Q000276:immunology*; D015658:HIV Infections / Q000276:immunology*; D015497:HIV-1 / Q000276:immunology*; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000235:genetics; D007074:Immunoglobulin G / Q000276:immunology*; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007147:Immunoglobulin Light Chains / Q000235:genetics; D007438:Introns; D007963:Leukocytes, Mononuclear; D009500:Neutralization Tests; D021382:Protein Sorting Signals / Q000235:genetics / Q000276:immunology; D011994:Recombinant Proteins / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D015704:CD4 Antigens; D015699:HIV Envelope Protein gp120; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D021382:Protein Sorting Signals; D011994:Recombinant Proteins