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The genes encoding the native-like trimers and HIV bnAbs were synthesized by GenScript. PBS pH 7.4 (Thermo Scientific, Catalog #10010-023) per well for assays detecting monoclonal and polyclonal antibodies, respectively, and incubated ON at 4°C. Human PGT121 Fab was used for trimers and non-nAbs capture (see Gene Synthesis and Protein Production), rabbit His-tag Antibody (GenScript, Catalog #A00174-40) was used for bnAbs capture. |
Elicitation of broadly neutralizing antibodies (bnAbs) is a primary HIV vaccine goal. Native-like trimers mimicking virion-associated spikes present nearly all bnAb epitopes and are therefore promising vaccine antigens. However, first generation native-like trimers expose epitopes for non-neutralizing antibodies (non-nAbs), which may hinder bnAb induction. We here employ computational and structure-guided design to develop improved native-like trimers that reduce exposure of non-nAb epitopes in the V3-loop and trimer base, minimize both CD4 reactivity and CD4-induced non-nAb epitope exposure, and increase thermal stability while maintaining bnAb antigenicity. In rabbit immunizations with native-like tri... More