Affinity maturation and characterization of a human monoclonal antibody against HIV-1 gp41.

Published

Journal Article

The human D5 monoclonal antibody binds to the highly conserved hydrophobic pocket on the N-terminal heptad repeat (NHR) trimer of HIV-1 gp41 and exhibits modest yet relatively broad neutralization activity. Both binding and neutralization depend on residues in the complementarity determining regions (CDRs) of the D5 IgG variable domains on heavy chain (VH) and light chain (VL). In an effort to increase neutralization activity to a wider range of HIV-1 strains, we have affinity matured the parental D5 scFv by randomizing selected residues in 5 of its 6 CDRs. The resulting scFv variants derived from four different CDR changes showed enhanced binding affinities to gp41 NHR mimetic (5-helix) which correlated to improved neutralization potencies by up to 8-fold. However, when converted to IgG1s, these D5 variants had up to a 12-fold reduction in neutralization potency over their corresponding scFvs despite their slightly enhanced in vitro binding affinities. Remarkably, D5 variant IgG1s bearing residue changes in CDRs that interact with epitope residues N-terminal to the hydrophobic pocket (such as VH CDR3 and VL CDR3) retained more neutralization potency than those containing residue changes in pocket-interacting CDRs (such as VH CDR2). These results provide compelling evidence for the existence of a steric block to an IgG that extends to the gp41 NHR hydrophobic pocket region, and can be a useful guide for developing therapeutic antibodies and vaccines circumventing this block.

Full Text

Duke Authors

Cited Authors

  • Montgomery, DL; Wang, Y-J; Hrin, R; Luftig, M; Su, B; Miller, MD; Wang, F; Haytko, P; Huang, L; Vitelli, S; Condra, J; Liu, X; Hampton, R; Carfi, A; Pessi, A; Bianchi, E; Joyce, J; Lloyd, C; Geleziunas, R; Bramhill, D; King, VM; Finnefrock, AC; Strohl, W; An, Z

Published Date

  • September 2009

Published In

Volume / Issue

  • 1 / 5

Start / End Page

  • 462 - 474

PubMed ID

  • 20065653

Pubmed Central ID

  • 20065653

Electronic International Standard Serial Number (EISSN)

  • 1942-0870

Digital Object Identifier (DOI)

  • 10.4161/mabs.1.5.9214

Language

  • eng

Conference Location

  • United States