Novel Quantitative, Comprehensive Proteomic Technology

描述：(申请人提供) 全球基因组计划产生了大量的信息， 摒弃新技术，催生了一种新型生物学 它试图从系统和定量的角度构建生物学知识 对构成细胞或组织的分子的分析。对…的期望 应用基因和蛋白质的全面和定量分析 癌症生物学很高。基因表达的mRNA和蛋白质的比较图谱 “正常”和癌细胞和组织有望发现新的标志物 癌症的类型、分期和分层的诊断、评估指标 治疗的成功和耐药性的出现，并导致 了解癌症的分子基础，从而找到新的治疗方法 目标。我们建议开发一种新的技术来定量、灵敏地 和全面分析人类细胞和 纸巾。它是基于络合物的定量、自动质谱学。 多肽混合物，由两个相组成。首先是引用的生成 给定细胞类型的多肽数据库，其中每个多肽都用 在数据库中唯一标识它的特征。第二个是 从临床或研究中获得的质谱学数据的相关性 样品与多肽数据库一起识别和量化蛋白质组分 细胞类型的。这项技术是：1)可自动化，2)兼容 基本上对任何蛋白质进行分析，3)健壮且易于标准化，4) 适应高吞吐量。这项技术将提供通用/健壮的 蛋白质综合定量分析的技术基础 正常细胞和癌细胞和组织的概况将被广泛 无障碍。这项新技术将得到验证并应用于前列腺 癌症研究。具体地说，我们将关联从 本项目开发的带基因的定量蛋白质图谱技术 通过基因芯片和Lynx治疗公司获得的大量表达数据 并行签名测序(MPSS)程序用于鉴定生物 区分惰性前列腺癌和侵袭性前列腺癌的决定因素 前列腺癌雄激素依赖性细胞生长的分子基础研究 细胞。该应用满足了对创新技术的要求 三个方面的发展：第一，这项技术探索了一种新的方法 蛋白质全谱的鉴定和定量 在人类细胞/组织中表达；第二，注释肽数据库(Apd) 因为前列腺组织将被公开提供，并作为必要的 实验室定量蛋白质表达图谱的参考文献 目前专门研究蛋白质组学，第三，研究将作为 学术/行业合作伙伴关系，确保快速实现商业可获得性 支持该技术的试剂/仪器。我们期待着拟议的 PRO公羊将为实现癌症的预期做出重大贡献 基因组学革命引发的研究。

DESCRIPTION: (provided by applicant) Global genome initiatives 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 systematic and quantitative analysis of the molecules that constitute a cell or tissue. Expectations for applying comprehensive and quantitative analyses of genes and proteins to cancer biology are high. Comparative profiling of mRNA & proteins expressed in "normal" and cancer cells and tissues is expected to discover new markers for diagnosis of type, stage and stratification of cancer, indicators to assess success of treatment and emergence of drug resistance, and lead to an understanding of the molecular basis of cancer and thus new therapeutic targets. We propose to develop a novel technology for quantitative, sensitive and comprehensive analysis of protein expression profiles in human cells and tissues. It is based on quantitative, automated mass spectrometry of complex peptide mixtures and consists of 2 phases. First is generation of a reference peptide database of a given cell type in which each peptide is annotated with characteristics that uniquely identify it in the database. Second is correlation of mass spectrometric data obtained from clinical or research samples with the peptide database to identify and quantify protein components of the cell type. The technology is: 1) automatable, 2) compatible with analysis of essentially any protein, 3) robust and easy to standardize and 4) amenable to high throughput. This technology will provide a general/robust technical basis for comprehensive and quantitative analysis of protein profiles of normal and cancer cells and tissues that will be widely accessible. The new technology will be validated and applied to prostate cancer research. Specifically, we will correlate the data obtained from the quantitative protein profiling technology developed in this program with gene expression data obtained via cDNA arrays and Lynx Therapeutic's massively parallel signature sequencing (MPSS) procedure to identify biologic determinants that differentiate indolent and aggressive prostate cancer and to determine the molecular basis of androgen dependent cell growth of prostate cells. The application satisfies the request for innovative technology development in 3 ways: First, the technology explores a new approach to identification and quantification of the comprehensive spectrum of proteins expressed in human cells/tissues; Second, the annotated peptide database (APD) for prostate tissue will be made available publicly and serve as a necessary reference for quantitative protein expression profiling in laboratories not currently specialized in proteomics and Third, research will be conducted as an academic/industrial partnership assuring rapid commercial accessibility of reagents/instrumentation supporting the technology. We anticipate the proposed pro ram will contribute significant towards fulfilling expectations for cancer research raised by the genomics revolution.