title: Dissociation rate of antibody-gp120 binding interactions is predictive of V3-mediated neutralization of HIV-1.
abstract: mAbs specific for the V3 loop of HIV-1 are capable of neutralizing laboratory strains of HIV-1 in vitro. In this report we use surface plasmon resonance and biosensor technology to demonstrate that the ability of V3-specific mAbs to neutralize HIV-1(MN) correlated with the dissociation rate constant of the homologous mAb-gp120 binding interaction. mAbs capable of binding diverse strains of gp120 with similar association rate constants exhibited marked differences in the dissociation rate. The dissociation rate, and not the association rate, was found to be predictive of the neutralization capacity of the mAb. Furthermore, synthetic peptides corresponding to the V3 loop of HIV-1(IIIB, MN) yielded quantitatively similar binding kinetic profiles abrogating the need for purified recombinant gp120 protein and potentially facilitating the screening of more diverse isolates. Biosensor immobilized V3 peptides were found to mimic their conformational structure in solution. This offers advantages to peptides studied by ELISA where some degree of denaturation and masking of epitopes can occur upon adsorption of peptides to plastic surfaces. The impact of amino acid substitutions within epitopes on subsequent mAb binding was dissected by observing alterations in dissociation rates. The technique provides rapid kinetic analyses of V3 Ab binding interactions with diverse V3 sequences, facilitating the efficient screening and selection of those most likely to possess the broadest and most potent HIV-1 neutralizing potentials.
authors.affiliation: Division of Retrovirology, Walter Reed Army Institute of Research, Rockville, MD 20850.
authors.forename: T C
authors.initials: TC
authors.lastname: VanCott
authors.forename: F R
authors.initials: FR
authors.lastname: Bethke
authors.forename: V R
authors.initials: VR
authors.lastname: Polonis
authors.forename: M K
authors.initials: MK
authors.lastname: Gorny
authors.forename: S
authors.initials: S
authors.lastname: Zolla-Pazner
authors.forename: R R
authors.initials: RR
authors.lastname: Redfield
authors.forename: D L
authors.initials: DL
authors.lastname: Birx
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000937:Antigen-Antibody Reactions; D000939:Epitopes; D015483:HIV Antibodies / Q000276:immunology / Q000378:metabolism*; D015699:HIV Envelope Protein gp120 / Q000276:immunology*; D006801:Humans; D007700:Kinetics; D008213:Lymphocyte Activation; D051379:Mice; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D010455:Peptides / Q000737:chemistry; D011485:Protein Binding; D013329:Structure-Activity Relationship
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D015483:HIV Antibodies; D015699:HIV Envelope Protein gp120; D010455:Peptides