ISOTOPE CODED AFFINITY TAGS FOR QUANTITATIVE PROTEOMICS

包括人类基因组计划在内的全球基因组计划产生了大量的信息，摒弃了新技术，并催化了一种新型生物学的出现，这种生物学试图通过对生物系统、途径和细胞的全球分析来建立生物学知识。在这个项目中，我们建议开发一种新的技术，用于定量和全局分析生物样品中的蛋白质表达谱，包括生化和亚细胞组分，生物液体，细胞和组织。因此，这项技术将把生物学研究的全球方法扩展到蛋白质的分析，蛋白质是普遍构成生物系统结构、功能和控制的分子。该技术的基础是一类新的试剂，我们称之为同位素编码亲和标签。在蛋白质分离后引入特定官能团的标签既可以作为分离标记蛋白质片段的配体，也可以作为质谱反卷积的定量代码。该方案具有三个显著特点，满足了创新技术发展的要求。首先，该技术探索了定量蛋白质分析的新方法，并有可能成为新兴的蛋白质组学领域的技术基础。其次，研究将由一个由学术/工业伙伴组成的多学科团队进行。这种伙伴关系预计将通过开发、商业化和支持所需的化学品、仪器和软件作为成熟的集成产品来加速技术的传播。第三，这项研究将在一个独特的研究环境中进行，即华盛顿大学新成立的定量系统生物学研究所。该研究所的结构体现了一项技术从诞生到融入生物研究项目和支持大规模应用的演变。该研究所还为学术/工业伙伴关系的蓬勃发展提供了跨学科的研究环境和设施。一旦发展起来，这种技术将成为生物学家试图从生物系统的功能、控制和机制方面解释基因组线性信息的重要工具。将该方法应用于癌症研究，将允许定量测量和识别区分特定癌细胞和正常细胞的分子。因此，这项技术将对分子诊断、癌症风险和预后的评估以及对癌症分子基础的理解做出重大贡献。

Global genome initiatives including the Human Genome Project have generated enormous amounts of information, spurned new technologies and catalyzed the emergence of a new type of biology which attempts to build biological knowledge from the global analysis of biological systems, pathways and cells. In this program we propose to develop a novel technology for the quantitative and global analysis of protein expression profiles in biological samples including biochemical and subcellular fractions, biological fluids, cells and tissues. Thus, this technology will extend the global approach to biological research to the analysis of proteins, the molecules that universally constitute the structure, function and control of biological systems. The basis of the technology is a new class of reagents we term isotope coded affinity tags. The tags which are introduced to specific functional groups in proteins post isolation serve as both, ligands for the isolation of tagged protein segments as well as a quantitative code which can be mass spectrometrically deconvoluted. The proposed program has three distinguishing features which satisfy the request for innovative technology development. First, the technology explores a new approach to quantitative protein analysis and has the potential to become the technical foundation of the emerging field of proteomics. Second, the research will be conducted by a multidisciplinary team constituted as an academic/industrial partnership. This partnership is expected to accelerate dissemination of the technology by developing, commercializing and supporting the required chemistries, instrumentation and software as mature, integrated products. Third, the research will be conducted in a unique research environment, the new Institute for Quantitative Systems Biology at the University of Washington. The structure of this institute embodies the evolution of a technology from its inception to the integration into biological research programs and to the support of large-scale applications. The Institute also provides the interdisciplinary research environment and the facilities required for the academic/industrial partnership to flourish. Once developed, such a technology will be an essential tool for biologists' attempts to interpret the linear information of genomes in terms of function, control and mechanisms of biological systems. Applied to cancer research the method will permit the quantitative measurement and identification of the molecules that distinguish a particular cancer cell from a normal cell. The technology will thus significantly contribute to the molecular diagnosis, the assessment of the cancer risk and prognosis, and the understanding of the molecular basis of cancer.