PROTEOMIC TECHNOLOGIES FOR CANCER RESEARCH

据估计，人类血清和尿液中含有5,000多种不同的蛋白质和肽，其中许多蛋白质和肽的浓度很低。 疾病相关的变化已被描述为大多数已被定量研究的不到5%，但很少有这些是有用的癌症标志物。因此，既迫切需要新的标记物，也需要探索大量潜在的标记物。 我们建议同时开发和测试技术，系统地和顺序地从正常人和癌症患者的血清和尿液中分离蛋白质，以发现有用的肿瘤标志物。 关键的分析工具是变性条件下的定量高分辨率二维电泳、用于蛋白质鉴定的质谱、快速循环免疫消减、制备和分析凝胶过滤、基于离子交换和疏水性的高分辨率色谱、等电聚焦和常规凝胶电泳。 结合起来，这些可以在很宽的动态范围内解析数千种蛋白质。 第一阶段将集中于证明新的蛋白质和一小组候选标记物确实可以使用这些技术以手动格式发现，而在第二阶段，每种方法将被机械化或自动化，以允许更高通量的癌症标记物发现。拟定商业应用：对癌症特有的蛋白质或蛋白质片段的鉴定将导致新的临床测试和用于成像小肿瘤块的方法的发展。 如果成功，这将彻底改变癌症的诊断和治疗。

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.