基于质谱技术的单细胞蛋白质组学

Mass Spectrometry Based Single-cell Proteomics

近年来,单细胞测序技术的发展极大地推动了单细胞基因组和转录组的研究。然而,直接关联单细胞生命过程的蛋白质组学研究则因技术进展缓慢而受限。随着样品前处理技术、色谱分离技术和质谱检测技术的进步,单细胞蛋白质组学(single-cell proteomics,SCP)的分析灵敏度显著提升。本综述深入回顾了SCP的发展历程及其在生命科学研究中的应用。在样品前处理方面,针对不同细胞的存在形式,研究者开发了基于声波激发、微流控芯片和激光切割等单细胞分选方法,并逐步从多步法前处理流程统一到一步法,降低了样品处理过程中的损失。在质谱技术方面,无标记定量(label-free quantification,LFQ)与基于同位素和同质素标记的方法都得到了充分的探索,各具技术优势与不足。在应用层面,SCP已在胚胎细胞早期发育、干细胞分化、肝脏组织空间异质性等方面揭示了新的生物学发现。最后,总结了当前SCP技术面临的挑战,包括检测通量、成本和数据分析的复杂性,并展望了其未来的发展方向,强调了SCP在基础研究和临床应用中的广阔前景。

In recent years,the development of single-cell sequencing technology has significantly advanced our understanding of single-cell genomics and transcriptomics.However,the study of proteomics,directly related to single-cell life processes,has been limited by slow technological progress.With advancements in sample preparation techniques and chromatography-mass spectrometry instruments,the analytical sensitivity of single-cell proteomics(SCP)has markedly improved.In this review,we thoroughly examine the development of SCP and its applications in life sciences.Regarding sample preparation,various methods such as gentle acoustic dispensing,microfluidic chips,and laser microdissection have been developed for single-cell sorting,gradually transitioning from multi-step to one-step processing,thereby reducing sample loss.In mass spectrometry,both label-free quantification and methods based on isotopic and isobaric labeling have been extensively explored,each with their own technical strengths and weaknesses.SCP has unveiled new biological insights in early embryonic cell development,stem cell differentiation,and spatial heterogeneity of liver tissues.Finally,we summarize the current challenges facing SCP technology,including detection throughput,cost,and data analysis complexity,while envisioning its future directions and emphasizing its broad potential in basic research and clinical applications.