HIV Neutralizing Antibody Assay
Monogram Biosciences’ HIV Neutralizing Antibody Assay is available to pharmaceutical and vaccine companies and researchers involved in HIV vaccine, therapeutic antibody research, and studies of the natural history of human immunodeficiency virus infection.
The assay is offered in three different formats: HITS, CURVES, and CURVES PLUS. HITS is a four-dilution format used for screening large numbers of plasmas; the CURVES format includes a 10-dilution curve that generates IC50s, IC80s, IC90s and IC95s; and the CURVES PLUS offering is the 10-dilution CURVES format plus the percent inhibition of virus at each antibody dilution.
Monogram maintains a large library of viral HIV-1 envelopes (gp160) populations from human plasma, etc, as well as single clones, including numerous non-subtype B viruses, which allow for virus panel customization. This library includes commonly available lab strain and repository viruses, as well as thousands of primary patient isolates. Clients may also send their own viruses (eg, a vaccine strain) for testing. Monogram Biosciences has recently developed a large panel of >400 single clones from worldwide clades. The antibody neutralization sensitivity, tropism, and gp160 sequences have been determined for all isolates. This panel represents currently circulating viruses from around the world and from more than 13 clades.
Monogram provides customized assays to answer targeted questions. A microneutralization assay was recently developed to screen B-cell culture supernatants and, in conjunction with a multi-institution team, Monogram’s screening assay identified >20 novel broadly neutralizing monoclonal antibodies.
Monogram is also very active in studying the coevolution of HIV envelope and the antibody response at various stages of infection. Monogram’s ability to clone virus envelope directly from any virus source, particularly patient plasma/serum samples, allows study of the evolving virus and antibody response in HIV-infected patients without the artificial selection inherent in culturing virus or selecting individual clones.
Examples of Assay Applications:
Characterization of Antibody Responses to Vaccination:
Gilbert P, Wang M, Wrin T, et al. Magnitude and breadth of a nonprotective neutralizing antibody response in an efficacy trial of a candidate HIV-1 gp120 vaccine. J Infec Dis. 2010;202(4):595-605. PMCID: PMC2946208
Du SX, Xu L, Zhang W, et al. A directed molecular evolution approach to improved immunogenicity of the HIV-1 envelope glycoprotein. PLoS One. 2011;6(6):e20927. PMCID: PMC3126809
Natural History Studies:
Richman DD, Wrin T, Little SJ, Petropoulos CJ. Rapid evolution of the neutralizing antibody response to HIV type 1 infection. Proc Natl Acad Sci USA. 2003 Apr 1;100(7):4144-4149.
Deeks SG, Schweighardt B, Wrin T, et al. Neutralizing antibody responses against autologous and heterologous viruses in acute versus chronic human immunodeficiency virus (HIV) infection: evidence for a constraint on the ability of HIV to completely evade neutralizing antibody responses. J Virol. 2006 Jun; 80(12):6155-6164.
Frost SD, Wrin T, Smith DM, et al. Neutralizing Antibody Responses Drive the Evolution of Human Immunodeficiency Virus Type 1 Envelope During Recent HIV Infection. Proc Natl Acad Sci U S A. 2005 Dec 20; 102(51):18514-18519.
Simek MD, Rida W, Priddy FH, et al. Human immunodeficiency virus type 1 elite neutralizers: individuals with broad and potent neutralizing activity identified by using a high-throughput neutralization assay together with an analytical selection algorithm. J Virol. 2009 Jul;83(14):7337-7348.
Discovery and Characterization of new MAbs:
Walker LM, Huber M, Doores KJ, et al. Broad neutralization coverage of HIV by multiple highly potent antibodies. Nature. 2011 Aug 17; 477(7365):466-470.
Walker LM, Phogat SK, Chan-Hui PY, et al. Broad and potent neutralizing antibodies from an African donor reveal a new HIV-1 vaccine target. Science. 2009 Oct 9; 326(5950):285-289.
O’Rourke SM, Schweighardt B, Scott WG, et al. Novel ring structure in the gp41 trimer of human immunodeficiency virus type 1 that modulates sensitivity and resistance to broadly neutralizing antibodies. J Virol. 2009 Aug; 83(15):7728-7738.
Binley JM1, Wrin T, Korber B, et al. Comprehensive cross-clade neutralization analysis of a panel of anti-human immunodeficiency virus type 1 monoclonal antibodies. J Virol. 2004 Dec; 78(23):13232-13252.