Department of Bioengineering, Northeastern University, Boston, MA, 02115, USA; Barnett Institute, Northeastern University, Boston, MA, 02115, USA; Department of Biology, Northeastern University, Boston, MA, 02115, USA.
Curr Opin Chem Biol. 2021 Feb;60:1-9. doi: 10.1016/j.cbpa.2020.04.018. Epub 2020 Jun 28.
Human physiology and pathology arise from the coordinated interactions of diverse single cells. However, analyzing single cells has been limited by the low sensitivity and throughput of analytical methods. DNA sequencing has recently made such analysis feasible for nucleic acids but single-cell protein analysis remains limited. Mass spectrometry is the most powerful method for protein analysis, but its application to single cells faces three major challenges: efficiently delivering proteins/peptides to mass spectrometry detectors, identifying their sequences, and scaling the analysis to many thousands of single cells. These challenges have motivated corresponding solutions, including SCoPE design multiplexing and clean, automated, and miniaturized sample preparation. Synergistically applied, these solutions enable quantifying thousands of proteins across many single cells and establish a solid foundation for further advances. Building upon this foundation, the SCoPE concept will enable analyzing subcellular organelles and posttranslational modifications, while increases in multiplexing capabilities will increase the throughput and decrease cost.
人类生理学和病理学源于多种单细胞的协调相互作用。然而，单细胞的分析受到分析方法灵敏度和通量低的限制。DNA 测序最近使得核酸的这种分析成为可能，但单细胞蛋白质分析仍然受到限制。质谱分析是蛋白质分析最强大的方法，但将其应用于单细胞面临三个主要挑战：将蛋白质/肽有效地递送到质谱检测器，识别其序列，并将分析扩展到数千个单细胞。这些挑战激发了相应的解决方案，包括 SCoPE 设计的多重化以及清洁、自动化和小型化的样品制备。协同应用这些解决方案，可以实现对数千个单细胞中数千种蛋白质的定量，并为进一步的进展奠定坚实的基础。在此基础上，SCoPE 概念将能够分析亚细胞细胞器和翻译后修饰，而多重化能力的提高将提高通量并降低成本。
