A Novel Diagnostic for Oncogenic HPV

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DESCRIPTION (provided by applicant): Arbor Vita proposes to detect and quantify oncogenic E6 proteins as a diagnostic test for Human Papilloma Virus (HPV) infection and cervical cancer. Oncogenic variants of HPVs have been correlated with cervical cancer at a frequency of 99.7%. Accumulated data unequivocally demonstrate that oncogenic E6 proteins are crucial for initiation and maintenance of cervical cell transformation and cancer progression, potentially representing a superior diagnostic marker. E6 proteins have not been successfully analyzed by standard methods. Research by Arbor Vita and others has shown that oncogenic E6 variants specifically bind to cellular PDZ domain proteins. Arbor Vita proposes to exploit this property as the basis of a simple test for the detection of oncogenic E6 variants. Specifically, Arbor Vita proposes to develop a PDZ domain-based oncogenic E6 detector that, as part of an ELISA-based test, selectively detects E6 oncoproteins: first, in vitro; secondly, in transfected cells; and thirdly, in HPV infected cancer cell lines. To accomplish these aims, Arbor Vita will utilize its proprietary in vitro assay. Tasks will include cloning, protein expression and purification, design of a ELISA based test system, and optimization of cell lysis conditions.