PROTEOMIC TECHNOLOGIES FOR CANCER RESEARCH

据估计，人的血清和尿液中含有5000多种不同的蛋白质和多肽，其中许多是微量的。在不到5%的定量研究中，大多数都描述了与疾病相关的变化，但很少有这些是有用的癌症标记物。因此，既迫切需要新的标记，也迫切需要探索大量潜在的标记。我们建议同时开发和测试从正常人和癌症患者的血清和尿液中系统地和顺序地分离蛋白质的技术，以发现有用的肿瘤标记物。主要的分析工具有变性条件下的定量高分辨双向电泳、蛋白质鉴定的质谱学、快速循环免疫吸附、制备和分析凝胶过滤、基于离子交换和疏水性的高分辨层析、等电聚焦和常规凝胶电泳。它们结合在一起，可以在很大的动态范围内分解数千种蛋白质。第一阶段将集中展示使用这些技术确实可以以手动形式发现新的蛋白质和一小部分候选标记，而在第二阶段，每种方法都将实现机械化或自动化，以实现更高吞吐量的癌症标记发现。拟议的商业应用：识别癌症特有的蛋白质或蛋白质片段将导致开发新的临床测试和对小肿瘤肿块进行成像的方法。如果成功，这将给癌症的诊断和治疗带来革命性的变化。

Human serum and urine are each estimated to contain over 5,000 different proteins and peptides, many in trace concentrations. Disease-associated changes have been described for most of the less than 5% which have been quantitatively studied, but very few of these are useful cancer markers. Thus there is both an urgent need for new markers, and a forest of potential markers to be explored. We proposed concurrent development and testing of technology for systematically and sequentially fractionating proteins from serum and urine from normal individuals and cancer patients to discover useful tumor markers. The key analytical tools are quantitative high resolution two-dimensional electrophoresis under denaturing conditions, mass spectroscopy for protein identification, rapid recycling immunosubtraction, preparative and analytical gel filtration, high resolution chromatography based on ion exchange and hydrophobicity, and isoelectric focusing and conventional gel electrophoresis. In combination these can resolve thousands of proteins over a wide dynamic range. Phase I will concentrate on the demonstration that new proteins and a small set of candidate markers can indeed be found using these techniques in a manual format, while in Phase II each method will be mechanized or automated to allow higher throughput cancer marker discovery. PROPOSED COMMERCIAL APPLICATION: Identification of proteins or protein fragments unique to cancer would lead to the development of new clinical tests and to methods for imaging small tumor masses. If successful, this would revolutionize the diagnosis and treatment of cancer.