Fine Molecular Specificity of Linear and Assembled Antibody Binding Sites in HIV-1 p24

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Abstract
A set of seven murine monoclonal antibodies were generated against a chemically synthesized 11-kDa 104-mer peptide covering the C-terminal residues 270-373 of the p24g.it; protein (HIV-IBRU strain). All monoclonal antibodies recognized HIV-LMN infected MOLT? cells by fluorescence und gave positive Western blot signals with viral gag peptides [p55 and/or p24), Oligopeptide binding regions were located with competitive enzyme-linked immunosorbent assays. Detailed epitope scanning analyses (the Geysen technique) were performed by serological testing of the monoclonal antibodies against 99 overlapping hexapeptides which corresponded to the entire 104-mer region. The antibodies bound to p24 peptide sequences located within the 275 2y3 and 351 36S regions. One antibody (LH 104-B) which reacted with residues 357-362 bound lo p55 alone. In contrast another antibody (LH 104–1). which recognized the residues 358–363. i.e. with five out of six residues in common with antibody LIIKM-B for its epitope region, reacted exclusively with p24. At least two of the antibodies (LH104-C and -A) which bound to p24 alone, apparently recognized conformational epitopes, They gave positive reactions with the regions 288—293/351–356 and 284–289/351–356, respectively. This work shows that chemical synthesis of large peptides is a viable alternative approach to immunochemical studies of viral proteins.