Ultrasensitive Proteome Profiling for 100 Living Cells by Direct Cell Injection, Online Digestion and Nano-LC-MS/MS Analysis
Ultrasensitive Proteome Profiling for 100 Living Cells by Direct Cell Injection, Online Digestion and Nano-LC-MS/MS Analysis
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通过直接细胞注射、在线消化和 Nano-LC-MS/MS 分析对 100 个活细胞进行超灵敏蛋白质组分析
DOI：
10.1021/acs.analchem.5b00808
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发表时间：
2015-07-07
影响因子：
7.4
通讯作者：
Zhang, Xiangmin
中科院分区：
文献类型：
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作者：
Chen, Qi;Yan, Guoquan;Zhang, Xiangmin
单细胞蛋白质组分析一直是一个令人兴奋的目标，因为它提供了关于细胞异质性和动态变化的关键信息。在这里，我们提出了一个集成的蛋白质组分析设备(IPad)100活细胞(iPad-100)，可能适合于单细胞分析。一旦细胞被培养，iPad-100就可以应用于注射100个活细胞，将活细胞转化为多肽，并以完全自动化的方式产生蛋白质鉴定结果。由于质谱学检测限的主要障碍，我们应用iPad-100分析了100个细胞的蛋白质组。通过三次重复运行，在DLD细胞蛋白质组中总共鉴定出813个蛋白质。基因本体论分析表明，来自不同细胞室的蛋白质，包括膜蛋白，都得到了很好的表达。IPad-100大大简化了采样过程，减少了样品损失，并防止了污染。结果，用iPad-100从100个细胞样本中鉴定出拷贝数低于1000的蛋白质，表明该方法的检测下限为200zmol。随着质谱学灵敏度的提高，iPad-100可能在不久的将来能够揭示单个细胞的丰富蛋白质组信息。
Single-cell proteome analysis has always been an exciting goal because it provides crucial information about cellular heterogeneity and dynamic change. Here we presented an integrated proteome analysis device (iPAD) for 100 living cells (iPAD-100) that might be suitable for single-cell analysis. Once cells were cultured, the iPAD-100 could be applied to inject 100 living cells, to transform the living cells into peptides, and to produce protein identification results with total automation. Due to the major obstacle for detection limit of mass spectrometry, we applied the iPAD-100 to analyze the proteome of 100 cells. In total, 813 proteins were identified in a DLD-cell proteome by three duplicate runs. Gene Ontology analysis revealed that proteins from different cellular compartments were well-represented, including membrane proteins. The iPAD-100 greatly simplified the sampling process, reduced sample loss, and prevented contamination. As a result, proteins whose copy numbers were lower than 1000 were identified from 100-cell samples with the iPAD-100, showing that a detection limit of 200 zmol was achieved. With increased sensitivity of mass spectrometry, the iPAD-100 may be able to reveal bountiful proteome information from a single cell in the near future.