Department of Pharmacology and Toxicology, UTMB Cancer Center, University of Texas Medical Branch, Galveston, Texas 77555, United States.
J Proteome Res. 2013 Jan 4;12(1):135-50. doi: 10.1021/pr3008607. Epub 2012 Dec 18.
A first research development progress report of the Chromosome 19 Consortium with members from Sweden, Norway, Spain, United States, China and India, a part of the Chromosome-centric Human Proteome Project (C-HPP) global initiative, is presented ( http://www.c-hpp.org ). From the chromosome 19 peptide-targeted library constituting 6159 peptides, a pilot study was conducted using a subset with 125 isotope-labeled peptides. We applied an annotation strategy with triple quadrupole, ESI-Qtrap, and MALDI mass spectrometry platforms, comparing the quality of data within and in between these instrumental set-ups. LC-MS conditions were outlined by multiplex assay developments, followed by MRM assay developments. SRM was applied to biobank samples, quantifying kallikrein 3 (prostate specific antigen) in plasma from prostate cancer patients. The antibody production has been initiated for more than 1200 genes from the entire chromosome 19, and the progress developments are presented. We developed a dedicated transcript microarray to serve as the mRNA identifier by screening cancer cell lines. NAPPA protein arrays were built to align with the transcript data with the Chromosome 19 NAPPA chip, dedicated to 90 proteins, as the first development delivery. We have introduced an IT-infrastructure utilizing a LIMS system that serves as the key interface for the research teams to share and explore data generated within the project. The cross-site data repository will form the basis for sample processing, including biological samples as well as patient samples from national Biobanks.
Chromosome 19 联盟的第一份研究进展报告,该联盟成员来自瑞典、挪威、西班牙、美国、中国和印度,是 Chromosome-centric Human Proteome Project (C-HPP) 全球倡议的一部分(http://www.c-hpp.org)。从构成 6159 个肽的染色体 19 肽靶向文库中,使用包含 125 个同位素标记肽的子集进行了试点研究。我们应用了具有三重四极杆、ESI-Qtrap 和 MALDI 质谱平台的注释策略,比较了这些仪器设置内和之间的数据质量。通过多重分析物检测开发了 LC-MS 条件,随后进行了 MRM 分析物检测开发。SRM 应用于生物银行样本,定量检测来自前列腺癌患者血浆中的激肽释放酶 3(前列腺特异性抗原)。已经为整个染色体 19 的 1200 多个基因启动了抗体生产,并且正在介绍进展情况。我们开发了一种专用的转录微阵列,通过筛选癌细胞系作为 mRNA 标识符。构建了 NAPPA 蛋白阵列,与 Chromosome 19 NAPPA 芯片上的转录数据对齐,该芯片专门用于 90 种蛋白质,作为第一个开发交付。我们引入了一个利用 LIMS 系统的 IT 基础设施,该系统作为研究团队共享和探索项目内生成的数据的关键接口。跨站点数据存储库将成为样本处理的基础,包括生物样本以及来自国家生物银行的患者样本。