A Novel Diagnostic for Oncogenic HPV

描述(申请人提供)：Arbor Vita建议检测和量化致癌的E6蛋白，作为人乳头瘤病毒(HPV)感染和宫颈癌的诊断测试。HPV的致癌变异与宫颈癌的相关性高达99.7%。积累的数据明确地表明，致癌的E6蛋白对宫颈细胞转化和癌症进展的启动和维持至关重要，可能是一种更好的诊断标记。E6蛋白还没有成功地用标准方法分析。Arbor Vita和其他人的研究表明，致癌的E6变体特异性地结合到细胞PDZ结构域蛋白上。Arbor Vita建议利用这一特性作为检测致癌E6变异的简单测试的基础。具体地说，Arbor Vita建议开发一种基于PDZ结构域的致癌E6检测器，作为基于ELISA测试的一部分，它选择性地检测E6癌蛋白：第一，在体外；第二，在转基因细胞中；第三，在HPV感染的癌细胞系中。为了实现这些目标，Arbor Vita将利用其专有的体外试验。任务将包括克隆、蛋白质表达和纯化、设计基于ELISA的测试系统以及优化细胞裂解条件。

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.