A Novel Diagnostic for Oncogenic HPV

描述（由申请人提供）：Arbor Vita建议检测和定量致癌E6蛋白，作为人乳头瘤病毒（HPV）感染和宫颈癌的诊断试验。HPV的致癌变异体与宫颈癌相关，频率为99.7%。累积的数据明确表明，致癌E6蛋白是至关重要的启动和维持宫颈细胞转化和癌症的进展，可能代表一个上级诊断标志物。E6蛋白还没有成功地通过标准方法进行分析。Arbor Vita和其他人的研究表明，致癌E6变体特异性结合细胞PDZ结构域蛋白。Arbor Vita建议利用这一特性作为检测致癌E6变体的简单测试的基础。具体来说，Arbor Vita提出开发一种基于PDZ结构域的致癌E6检测器，作为基于ELISA的测试的一部分，选择性地检测E6癌蛋白：首先，在体外;其次，在转染的细胞中;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，蛋白表达和纯化，基于ELISA的测试系统的设计，以及细胞裂解条件的优化。

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.