Zhang Zhaorui, Wu Si, Stenoien David L, Paša-Tolić Ljiljana
Environmental Molecular Sciences Laboratory.
Annu Rev Anal Chem (Palo Alto Calif). 2014;7:427-54. doi: 10.1146/annurev-anchem-071213-020216.
Mass spectrometry (MS)-based high-throughput proteomics is the core technique for large-scale protein characterization. Due to the extreme complexity of proteomes, sophisticated separation techniques and advanced MS instrumentation have been developed to extend coverage and enhance dynamic range and sensitivity. In this review, we discuss the separation and prefractionation techniques applied for large-scale analysis in both bottom-up (i.e., peptide-level) and top-down (i.e., protein-level) proteomics. Different approaches for quantifying peptides or intact proteins, including label-free and stable-isotope-labeling strategies, are also discussed. In addition, we present a brief overview of different types of mass analyzers and fragmentation techniques as well as selected emerging techniques.
基于质谱（MS）的高通量蛋白质组学是大规模蛋白质表征的核心技术。由于蛋白质组极其复杂，已开发出复杂的分离技术和先进的质谱仪器，以扩大覆盖范围、提高动态范围和灵敏度。在本综述中，我们讨论了在自下而上（即肽水平）和自上而下（即蛋白质水平）蛋白质组学中用于大规模分析的分离和预分级技术。还讨论了定量肽或完整蛋白质的不同方法，包括无标记和稳定同位素标记策略。此外，我们简要概述了不同类型的质量分析器和裂解技术以及选定的新兴技术。
