Zhou Weidong, Petricoin Emanuel F, Longo Caterina
Center for Applied Proteomics and Molecular Medicine, George Mason University, 10920 George Mason Circle, MS1A9, Manassas, VA, 20110, USA.
Dermatology and Skin Cancer Unit, Arcispedale S Maria Nuova IRCCS, Reggio Emilia, Italy.
Methods Mol Biol. 2017;1606:297-311. doi: 10.1007/978-1-4939-6990-6_19.
The discovery of candidate biomarkers within the entire proteome is one of the most important and challenging goals in proteomic research. Mass spectrometry-based proteomics is a modern and promising technology for semiquantitative and qualitative assessment of proteins, enabling protein sequencing and identification with exquisite accuracy and sensitivity. For mass spectrometry analysis, protein extractions from tissues or body fluids and subsequent protein fractionation represent an important and unavoidable step in the workflow for biomarker discovery. Following extraction of proteins, the protein mixture must be digested, reduced, alkylated, and cleaned up prior to mass spectrometry. The aim of our chapter is to provide comprehensible and practical lab procedures for sample digestion, protein fractionation, and subsequent mass spectrometry analysis.
在整个蛋白质组中发现候选生物标志物是蛋白质组学研究中最重要且最具挑战性的目标之一。基于质谱的蛋白质组学是一种用于蛋白质半定量和定性评估的现代且有前景的技术，能够以极高的准确性和灵敏度进行蛋白质测序和鉴定。对于质谱分析而言，从组织或体液中提取蛋白质以及随后的蛋白质分级分离是生物标志物发现工作流程中重要且不可避免的步骤。蛋白质提取后，在进行质谱分析之前，必须对蛋白质混合物进行消化、还原、烷基化和净化处理。本章的目的是提供关于样品消化、蛋白质分级分离以及后续质谱分析的易懂且实用的实验室操作程序。
