High throughput proteome screening for biomarker detection
High throughput proteome screening for biomarker detection
复制标题
DOI：
10.1074/mcp.m400161-mcp200
复制
发表时间：
2005-02-01
影响因子：
7
通讯作者：
Aebersold, R
中科院分区：
文献类型：
--
作者：
Pan, S;Zhang, H;Aebersold, R
基于质谱的定量蛋白质组学已成为生物学和临床研究的重要组成部分。目前的方法，虽然高度发达和强大的，是达不到他们的目标，常规分析整个蛋白质组，主要是因为丰富的蛋白质组信息积累从以前的研究是不用于规划或解释目前的实验。这种情况的后果是，在每一个蛋白质组学实验中，蛋白质组被重新发现。在这份报告中，我们描述了一种定量蛋白质组学的方法，建立在广泛的蛋白质组学的先验知识和平台的方法的实施。该方法基于同位素标记的参考肽的选择和化学合成，其唯一地识别特定蛋白质，并且将一组这样的肽添加到由来自所讨论的蛋白质组的胰蛋白酶肽组成的样品混合物中。该平台包括一个肽分离模块，用于从MALDI质谱仪样品板上的组合肽样品生成有序肽阵列，一个高通量MALDI-TOF/TOF质谱仪，以及一套用于选择性分析靶肽和解释结果的软件工具。将该方法应用于人血清蛋白质组的分析，我们证明了使用基于质谱的蛋白质组学作为复杂系统中目标蛋白质的检测和定量的高通量筛选技术的可行性。
Mass spectrometry-based quantitative proteomics has become an important component of biological and clinical research. Current methods, while highly developed and powerful, are falling short of their goal of routinely analyzing whole proteomes mainly because the wealth of proteomic information accumulated from prior studies is not used for the planning or interpretation of present experiments. The consequence of this situation is that in every proteomic experiment the proteome is rediscovered. In this report we describe an approach for quantitative proteomics that builds on the extensive prior knowledge of proteomes and a platform for the implementation of the method. The method is based on the selection and chemical synthesis of isotopically labeled reference peptides that uniquely identify a particular protein and the addition of a panel of such peptides to the sample mixture consisting of tryptic peptides from the proteome in question. The platform consists of a peptide separation module for the generation of ordered peptide arrays from the combined peptide sample on the sample plate of a MALDI mass spectrometer, a high throughput MALDI-TOF/TOF mass spectrometer, and a suite of software tools for the selective analysis of the targeted peptides and the interpretation of the results. Applying the method to the analysis of the human blood serum proteome we demonstrate the feasibility of using mass spectrometry- based proteomics as a high throughput screening technology for the detection and quantification of targeted proteins in a complex system.