Methods for the detection of HIV-1-specific antibodies employing GP41 polypeptides
Inventors
Golding, Hana • Khurana, Surender
Assignees
US Department of Health and Human Services
Publication Number
US-8722324-B2
Publication Date
2014-05-13
Expiration Date
2025-09-02
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Abstract
This invention relates to compositions and methods or the detection of human immunodeficiency virus-1 (HIV-1) infection by conducting an immunoassay comprising the steps of: (a) contacting a biological sample containing HIV-1 antibody with a peptide, having an epitope, of one or more of SEQ ID 49-56 to form a peptide-anti-HIV-1 antibody complex; (b) contacting the formed complex with an anti-HIV-1 antibody binding molecule to permit the anti-HIV-1 antibody binding molecule to bind to the anti-HIV-1 antibody of the formed peptide-anti-HIV-1 antibody complex and form an extended complex that is immobilized on a solid support; (c) removing unbound anti-HIV-1 antibody and anti-HIV-1 antibody binding molecule from the extended complex; and (d) determining the presence or concentration of the anti-HIV-1 antibody in the biological sample.
Core Innovation
This invention relates to compositions and methods for the detection of human immunodeficiency virus-1 (HIV-1) infection by conducting an immunoassay. The method involves contacting a biological sample comprising anti-HIV-1 antibody with a peptide having an epitope of one or more selected sequences (e.g., SEQ ID NOs 49-56), forming a peptide-antibody complex. This complex is then contacted with an anti-HIV-1 antibody binding molecule to form an extended complex immobilized on a solid support, followed by removal of unbound components and determination of the antibody presence or concentration.
The problem addressed is the difficulty in distinguishing between antibodies generated by HIV infection and those generated by HIV vaccines, particularly in vaccine recipients. Current licensed HIV serodetection assays cannot differentiate between vaccine-induced and infection-induced antibody responses. This leads to vaccine recipients' sera reacting indistinguishably from infected individuals, complicating early detection of breakthrough infections during vaccine trials, excluding vaccinated individuals from blood donation, and causing socio-economic harms related to false positive HIV diagnoses.
The invention identifies new HIV-1 and HIV-2 epitopes that are broadly reactive with early serum samples from individuals infected with HIV strains from all clades, do not contain protective or cytotoxic epitopes, and are not included in current or future HIV vaccine candidates. Using gene-fragment phage display libraries from whole HIV genomes, the invention constructs differential enzyme-immunoassays capable of distinguishing infection-induced antibodies from vaccine-induced reactivities. Peptides derived from conserved regions of HIV-1 Gag p6 and gp41, as well as from HIV-2 Gag p6 and Env-gp36, are central to these assays.
Claims Coverage
The patent contains multiple independent claims covering various methods and compositions for detecting anti-HIV antibodies using specific peptides.
Method for detecting anti-HIV-1 antibodies using non-immobilized peptides and immobilized binding molecules
A method comprising: (a) contacting a biological sample with a non-immobilized peptide having an epitope recognized by anti-HIV-1 antibody to form a peptide-anti-HIV-1 antibody complex; (b) contacting said complex with an anti-HIV-1 antibody binding molecule immobilized on a solid support to form an extended complex immobilized on the solid support; (c) removing unbound anti-HIV-1 antibody and/or binding molecules; and (d) determining the presence or concentration of anti-HIV-1 antibody by detecting the extended complex, using ELISA or immunochromatographic assays; wherein the epitope is present on peptides with sequences selected from SEQ ID NOs 50-52.
Use of peptides or proteins comprising specific HIV-1 epitopes in detection methods
Employing peptides or proteins having amino acid sequences SEQ ID NOs: 50, 51, or 52 as antigenic components to recognize anti-HIV-1 antibodies in immunoassays.
Immunological complexes comprising peptides and anti-HIV-1 antibodies with anti-HIV antibody binding molecules
Formation of an immunological complex comprising a peptide bound to an anti-HIV-1 antibody, which is further bound to an anti-HIV antibody binding molecule, wherein the peptide epitope is from specific sequences like SEQ ID NOs 1-11, 49-56, 90, or 141.
Kits for detection of anti-HIV-1 antibodies
Kits comprising a hollow casing with a multilayer filter system and porous carriers, where the first carrier contains a non-immobilized, labeled peptide or protein comprising specific epitopes and the second carrier contains an immobilized, unlabeled antibody that binds human IgG, intended for detection of anti-HIV-1 antibodies.
Methods for detecting anti-HIV-2 antibodies with specific peptides
Methods mimicking the anti-HIV-1 antibody detection approach, using peptides with epitopes present in SEQ ID NOs 101-102, for detecting or measuring anti-HIV-2 antibodies.
Use of epitope sets comprising HIV-1 Gag p6 and gp41 terminal region epitopes
Conducting immunoassays using epitope sets consisting essentially of HIV-1 Gag p6 epitopes, HIV-1 gp41 terminal region epitopes, or combinations thereof, for detecting anti-HIV-1 antibodies.
Use of epitope sets comprising HIV-2 Gag p6 and Env-gp36 epitopes
Conducting immunoassays using epitope sets consisting essentially of HIV-2 Gag p6 epitopes, HIV-2 Env-gp36 epitopes, or combinations thereof, for detecting anti-HIV-2 antibodies.
The independent claims cover methods of detecting anti-HIV-1 and anti-HIV-2 antibodies by immunoassays utilizing peptides with defined epitopes (SEQ ID NOs: 1-11, 49-56, 90, 101-102, 141), formulations of immunological complexes, specific use of peptides in such methods, and kits for such detection. The inventive features focus on employing selected conserved epitopes not present in vaccines, allowing differentiation between vaccine-induced and infection-induced antibodies.
Stated Advantages
Allows differentiation of HIV infection-induced antibodies from vaccine-induced antibodies, enabling early detection of breakthrough infections in vaccine recipients.
Provides high sensitivity and specificity across diverse HIV clades and subtypes, including early infection.
Reduces false positives in vaccinated individuals, thereby allowing accurate diagnosis and minimizing socio-economic harms such as exclusion from blood donation, insurance, employment, and travel.
Enables inexpensive, high-throughput assays suitable for clinical and blood donation screening.
Detects intercurrent HIV infections earlier or concurrent with licensed tests, improving disease management during vaccine trials.
Documented Applications
Use in immunoassays for the detection of anti-HIV-1 and anti-HIV-2 antibodies in biological samples from humans.
Differentiation between vaccine-induced antibodies and infection-induced antibodies during HIV vaccine clinical trials.
Screening for early HIV-1 and HIV-2 infection diagnosis, including acute seroconversion detection.
Monitoring of individuals during vaccination campaigns to identify breakthrough infections.
Blood and plasma donor screening to prevent inclusion of infected individuals while allowing vaccinated but uninfected individuals to donate.
Use in kits comprising solid supports and porous carriers for ELISA and immunochromatographic assays.
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