Antibody conjugation approach enhances breadth and potency of neutralization of anti-HIV-1 antibodies and CD4-IgG

J Virol. 2013 May;87(9):4985-93. doi: 10.1128/JVI.03146-12. Epub 2013 Feb 20.

Abstract

Broadly neutralizing antibodies PG9 and PG16 effectively neutralize 70 to 80% of circulating HIV-1 isolates. In this study, the neutralization abilities of PG9 and PG16 were further enhanced by bioconjugation with aplaviroc, a small-molecule inhibitor of virus entry into host cells. A novel air-stable diazonium hexafluorophosphate reagent that allows for rapid, tyrosine-selective functionalization of proteins and antibodies under mild conditions was used to prepare a series of aplaviroc-conjugated antibodies, including b12, 2G12, PG9, PG16, and CD4-IgG. The conjugated antibodies blocked HIV-1 entry through two mechanisms: by binding to the virus itself and by blocking the CCR5 receptor on host cells. Chemical modification did not significantly alter the potency of the parent antibodies against nonresistant HIV-1 strains. Conjugation did not alter the pharmacokinetics of a model IgG in blood. The PG9-aplaviroc conjugate was tested against a panel of 117 HIV-1 strains and was found to neutralize 100% of the viruses. PG9-aplaviroc conjugate IC50s were lower than those of PG9 in neutralization studies of 36 of the 117 HIV-1 strains. These results support this new approach to bispecific antibodies and offer a potential new strategy for combining HIV-1 therapies.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Neutralizing / chemistry
  • Antibodies, Neutralizing / immunology*
  • Antibodies, Neutralizing / pharmacology
  • Benzoates / chemistry
  • Benzoates / pharmacology
  • CD4 Immunoadhesins / chemistry
  • CD4 Immunoadhesins / immunology*
  • CD4 Immunoadhesins / pharmacology
  • Cell Line
  • Diketopiperazines
  • HIV Antibodies / chemistry
  • HIV Antibodies / immunology*
  • HIV Antibodies / pharmacology
  • HIV Infections / drug therapy
  • HIV Infections / immunology*
  • HIV Infections / virology
  • HIV-1 / drug effects
  • HIV-1 / immunology*
  • HIV-1 / physiology
  • Humans
  • Immunoconjugates / chemistry
  • Immunoconjugates / immunology*
  • Immunoconjugates / pharmacology
  • Neutralization Tests
  • Piperazines / chemistry
  • Piperazines / pharmacology
  • Spiro Compounds / chemistry
  • Spiro Compounds / pharmacology
  • Virus Internalization / drug effects

Substances

  • Antibodies, Neutralizing
  • Benzoates
  • CD4 Immunoadhesins
  • Diketopiperazines
  • HIV Antibodies
  • Immunoconjugates
  • Piperazines
  • Spiro Compounds
  • aplaviroc