Reverse Immunology Approach to Define a New HIV-gp41-Neutralizing Epitope

Figure 3

Phage-mimotope selection and reactivity of the anti-W614A-3S mAbs. A phage library of decapeptides was screened with the F8 mAb (a). After three rounds of selection in the first biopanning, the mim_F8-1 phagotope represented 25% of the selected clones. In a second experiment of biopanning, three phagotopes were identified at the third round of selection. Mim_F8-2 represented 42% of the selected clones and shared a five-residue motif with those of Mim_F8-1. (b) Binding properties of the selected phagotopes were tested by ELISA on immobilized anti-W614A-3S mAbs. Binding activity is expressed as mean OD at 450 nm of duplicate wells and bars indicate SD. These data are representative of three independent experiments. (c) Competitive binding ELISA of Mim_F8-1 on the immobilized F8 mAb with indicated concentrations of BSA-W614A-3S (black bars). BSA-3S-scr functioned as negative control (white bars). Results are expressed as percentage of binding of the Mim_F8-1 without competitor (100%). Values are the means of two different experiments with two replicated measurements, and bars indicate SEM. , according to Mann–Whitney test.

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