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HIV A12

A12 is a neutralizing VHH (single domain antibody) Ab devoid of light chains. It was isolated from sera from llamas, who produce immunoglobulins devoid of light chains, immunized with gp120 of HIV-1 CRF07_BC primary isolate CN54, following panning of phage libraries expressing VHH repertoire and a competitive elution with soluble CD4. It was hypothesized that the small size of the VHH, in combination with their protruding CDR3 loops, and their preference for cleft recognition and binding into active sites, may allow for recognition of conserved motifs on gp120 that are occluded from conventional Abs. A12 was able to neutralize HIV-1 primary isolates of subtypes B, C and CRF07_BC, but not subtypes A, D, and A/G. Compared to MAb b12, which neutralized 54% of viruses tested, A12 neutralized 42% of the viruses, but it neutralized a different spectrum of the viruses than b12. A12 showed high affinity binding to IIIB gp120, and inhibited binding of sCD4 to IIIB gp120 and 92UG037 gp140 in a dose-dependent manner. A12 was found to compete with b12 for binding to gp120, and also with MAbs 654-D and GP68, indicating that its epitope overlaps the CD4bs. There was some inhibition observed of A12-gp120 binding by 2G12, 17b, and 447-52D, while 4E10 did not inhibit A12-gp120 binding. A12 was also able to inhibit binding of the other two VHH Abs isolated in this study, D7 and C8, indicating that their epitopes overlap.

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