10.1021/bi0003691.s001 Edelmira Cabezas Edelmira Cabezas Meng Wang Meng Wang Paul W. H. I. Parren Paul W. H. I. Parren Robyn L. Stanfield Robyn L. Stanfield Arnold C. Satterthwait Arnold C. Satterthwait A Structure-Based Approach to a Synthetic Vaccine for HIV-1<sup>†</sup> American Chemical Society 2000 loop 5 PAbs protein neutralization sites MN envelope glycoprotein NMR V 3 MAb 58.2 GPGR V 3 peptide approach JHIGPGR antibody NH CH HIV loop 5 2000-11-01 00:00:00 Journal contribution https://acs.figshare.com/articles/journal_contribution/A_Structure-Based_Approach_to_a_Synthetic_Vaccine_for_HIV-1_sup_sup_/3584418 The generation of neutralizing antibodies by peptide immunization is dependent on achieving conformational compatibility between antibodies and native protein. Consequently, approaches are needed for developing conformational mimics of protein neutralization sites. We replace putative main-chain hydrogen bonds (NH → OCRNH) with a hydrazone link (N−NCH−CH<sub>2</sub>CH<sub>2</sub>) and scan constrained peptides for fit with neutralizing monoclonal antibodies (MAbs). To explore this approach, a V3 MAb 58.2 that potently neutralizes T-cell lab-adapted HIV-1<sub>MN</sub> was used to identify a cyclic peptide, [JHIGPGR(Aib)F(d-Ala)GZ]G-NH<sub>2</sub> (loop 5), that binds with >1000-fold higher affinity than the unconstrained peptide. NMR structural studies suggested that loop 5 stabilized β-turns at GPGR and R(Aib)F(d-Ala) in aqueous solvent implying considerable conformational mimicry of a Fab 58.2 bound V3 peptide determined by X-ray crystallography [Stanfield, R. L. et al. (1999) <i>Structure</i> <i>142</i>, 131−142]. Rabbit polyclonal antibodies (PAbs) generated to loop 5 but not to the corresponding uncyclized peptide bound the HIV-1<sub>MN</sub> envelope glycoprotein, gp120. When individual rabbit antisera were scanned with linear and cyclic peptides, further animal-to-animal differences in antibody populations were characterized. Loop 5 PAbs that most closely mimicked MAb 58.2 neutralized HIV-1<sub>MN</sub> with similar potency. These results demonstrate the remarkable effect that conformation can have on peptide affinity and immunogenicity and identify an approach that can be used to achieve these results. The implications for synthetic vaccine and HIV-1 vaccine research are discussed.