高通量蛋白质组学分析研究进展
摘要
<p id=C3>Proteomic analysis aims at characterizing proteins on a large scale, including their relative abundance, post-translational modifications, protein-protein interactions and so on. Proteomic profiling helps to elucidate the mechanisms of disease occurrence and to discover new diagnostic markers and therapeutic targets. Mass spectrometry (MS)-based proteomic technologies have advanced to allow comprehensive qualitative and quantitative proteome profiling across a myriad proteins in cells and tissues. High-throughput proteomics is the core technique for large-scale protein characterization. With the increased demand for large cohort proteomic analysis in the biomedical research field, high-throughput proteomic analysis has become a critical issue that needs to be urgently addressed. The standard shotgun proteomic workflow comprises four steps, including sample preparation, peptide separation, MS acquisition, and data analysis. Advances in these four steps have contributed to the development of high-throughput proteomics. In this review, we aimed at summarizing the current information on the state-of-the-art development of high-throughput proteomic analysis, mainly including the following topics: (1) High-throughput, automatic proteomic sample preparation methods based on liquid-handling workstations. The automation of the proteomic sample preparation steps is essential for high-throughput proteomic analysis, which will significantly reduce variation of manual operation and sample loss by multistep sample processing. The commercial liquid handling workstations, including King Fisher<sup>TM</sup> Flex, Agilent Bravo, AssayMAP Bravo, and Biomek<sup>®</sup> NX<sup>P</sup>, perform the handling steps of 96- or 384-channel microplate formats using a mechanical arm that increases the throughput and robustness of sample preparation. (2) High-throughput proteomic detection methods based on microliter-flow-rate liquid chromatography coupled with mass spectrometry (micro-