Method for the detection of HIV-1 antibodies utilizing a peptide containing a novel gp41 epitope
Inventors
Golding, Hana • Khurana, Surender
Assignees
US Department of Health and Human Services
Publication Number
US-9121855-B2
Publication Date
2015-09-01
Expiration Date
2025-09-02
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Abstract
This invention concerns methods for detecting the presence of an anti-HIV-1 antibody in a biological sample, the method comprising conducting an immunoassay comprising: (a) contacting the biological sample with at least one epitope that is recognized by the anti-HIV-1 antibody, wherein the contacting being under conditions sufficient to permit the anti-HIV-1 antibody if present in the sample to bind to the epitope and form an epitope-anti-HIV-1 antibody complex; (b) contacting the formed epitope-anti-HIV-1 antibody complex with an anti-HIV-1 antibody binding molecule, the contacting being under conditions sufficient to permit the anti- HIV-1 antibody binding molecule to bind to anti-HIV-1 antibody of the formed epitope-anti-HIV-1 antibody complex and form an extended complex; and (c) determining the presence or concentration of the anti-HIV-1 antibody in the biological sample by determining the presence or concentration of the formed extended complex; the epitope being present on a peptide comprising SEQ ID No. 55.
Core Innovation
This invention concerns compositions and methods for the detection of immunodeficiency virus infection, especially human immunodeficiency virus-1 (HIV-1) and human immunodeficiency virus-2 (HIV-2) infection. It particularly addresses the challenge in HIV vaccine recipients whose sera may contain vaccine-generated anti-HIV antibodies, creating difficulty in distinguishing these antibodies from those present due to true infection.
The invention identifies new HIV-1 and HIV-2 epitopes which are broadly reactive with early serum samples from individuals infected with HIV virus strains from all clades, do not contain protective antibody or cytotoxic epitopes, and can be easily excluded from current and future HIV-1 vaccine candidates. This is achieved by constructing gene-fragment phage display libraries from the whole HIV-1 genome to discover such epitopes and develop differential enzyme-immunoassays capable of distinguishing infection-induced anti-HIV antibodies from vaccine-induced reactivities.
The identified HIV-1 epitopes, especially from the gag p6 and gp41 terminal region, and HIV-2 epitopes from GAG p6 and Env-gp36, enable the development of sensitive and specific immunoassays. These immunoassays utilize peptides containing such epitopes for detecting anti-HIV antibodies in biological samples, thus providing a means to discern between antibodies arising from infection and those generated by vaccination. The compositions include synthetic peptides with specific sequences (e.g., SEQ ID NO:3, SEQ ID NO:50, SEQ ID NO:55) and methods including ELISA and immunochromatographic assays.
Claims Coverage
The patent includes two independent claims directed to methods for detecting anti-HIV-1 antibodies in biological samples using immunoassays with specific peptides containing novel gp41 epitopes.
Method for detecting anti-HIV-1 antibodies using gp41 peptide epitope
A method for conducting an immunoassay comprising: (a) contacting a biological sample with at least one epitope present on a peptide consisting essentially of SEQ ID No. 55 to bind anti-HIV-1 antibodies if present; (b) contacting the formed epitope-antibody complex with an anti-HIV-1 antibody binding molecule to form an extended complex; and (c) determining the presence or concentration of anti-HIV-1 antibodies by detecting the extended complex.
Immunoassay format incorporating ELISA and immunochromatography
The method wherein the immunoassay is an ELISA with the peptide immobilized on a solid support coated with milk, and anti-HIV-1 antibody binding molecules are selected from anti-human IgG+IgM-Fc, anti-human IgG-Fc, anti-human IgG+IgM, and anti-human IgG conjugated to an enzyme. Alternatively, the immunoassay is an immunochromatographic assay using two porous carriers, the first containing labeled non-immobilized peptide and the second containing immobilized unlabeled anti-HIV-1 antibody binding molecule, whereby the complex moves between carriers and is detected.
The independent claims cover novel immunoassay methods for detecting anti-HIV-1 antibodies in biological samples by employing specific novel gp41 peptide epitopes and corresponding antibody binding molecules in ELISA and immunochromatographic formats, enabling detection of antibodies indicative of HIV-1 infection with specificity.
Stated Advantages
Provides a simple and inexpensive assay that does not score uninfected vaccine recipients as positive, improving specificity for distinguishing vaccine-induced antibodies from infection.
Allows early detection of breakthrough HIV infections during vaccine trials, critical for appropriate clinical management and trial integrity.
Enables differential diagnosis to minimize social and economic harms to vaccine recipients erroneously diagnosed as infected.
Applicable worldwide for detecting diverse HIV-1 clades and subtypes, supporting global vaccine trials across different populations.
The immunoassay is high-throughput, low cost, and suitable for clinical and blood/plasma donation settings.
Documented Applications
Use in clinical diagnostic assays to detect the presence or concentration of anti-HIV-1 antibodies in biological samples such as serum or plasma.
Application as differential assays in HIV vaccine clinical trials to distinguish between antibodies generated by vaccination and those resulting from true HIV-1 infection.
Screening in blood and plasma donation centers to avoid exclusion of healthy vaccine recipients while ensuring infected individuals are identified.
Monitoring of early HIV-1 infections in vaccine recipients to enable timely clinical intervention.
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