Abstract 7086: Serological assays for early detection of HPV-associated oropharyngeal cancer

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Abstract Research Use Only (RUO) sample-sparing MULTI-ARRAY serology immunoassays have been developed for human antibodies to several HPV-16 early antigens. These assays are intended to support research of oropharyngeal cancer (OPC) linked to the human papillomavirus (HPV).The HPV virus can infect the mouth and throat and cause cancers of the oropharynx, i.e. the back of the throat, including the base of the tongue and tonsils. HPV-related cancer incidence is increasing dramatically among men and is unlikely to decrease in the foreseeable future, despite vaccination efforts. As a result, this may become one of the most common cancers in middle-aged men in the United States by 2045. The HPV16 genome consists of six early genes (E1, E2, E4, E5, E6, and E7) and two late genes (L1 and L2) that constitute the viral capsid. Antibodies to early HPV-16 antigens are commonly present in those with OPC but are detected in only approximately 1% of the general population. These antibodies are present many years before cancer develops; thus, they are ideal biomarkers for early detection of OPC.Immunoassays for antibodies against HPV-16 early antigens E1, E2, E6, and E7 were developed in a high-throughput multiplexed format using electrochemiluminescence (ECL) detection, and MULTI-ARRAY 10-spot 96-well plates. Each well has an array presenting recombinant antigens manufactured by Meso Scale Diagnostics, LLC. The assay uses 25 uL of 2, 500x diluted serum or plasma. The assay format is simple: diluted sample is incubated in the well with the antigen array followed by a wash and detection of bound anti-HPV antibodies using an anti-human IgG antibody labeled with the SULFO-TAG ECL label. Assay performance was evaluated with approximately 200 commercially sourced samples from apparently healthy individuals and 14 samples from individuals known to be positive for at least one HPV-16 early antigen using an established reference method (Programmable Protein (RAPID) ELISA, Anderson 2015). There was excellent agreement between the two methods.This high-throughput MULTI-ARRAY assay may be useful in research applications requiring screening of a large number of samples for HPV-16 antibodies.For Research Use Only. Not for use in diagnostic procedures. Citation Format: Mingyue Wang, Jermaine Brown, John C. Smith, Leonid Dzantiev, Anu Mathew, Martin Stengelin, George Sigal, Padhma Yuvaraj, Karen S. Anderson, Jacob Wohlstadter. Serological assays for early detection of HPV-associated oropharyngeal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 7086.

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