College of Pharmacy, Gachon University, Incheon 21936, Korea.
Int J Mol Sci. 2020 Feb 23;21(4):1524. doi: 10.3390/ijms21041524.
Proteomics is a large-scale study of proteins, aiming at the description and characterization of all expressed proteins in biological systems. The expressed proteins are typically highly complex and large in abundance range. To fulfill high accuracy and sensitivity of proteome analysis, the hybrid platforms of multidimensional (MD) separations and mass spectrometry have provided the most powerful solution. Multidimensional separations provide enhanced peak capacity and reduce sample complexity, which enables mass spectrometry to analyze more proteins with high sensitivity. Although two-dimensional (2D) separations have been widely used since the early period of proteomics, three-dimensional (3D) separation was barely used by low reproducibility of separation, increased analysis time in mass spectrometry. With developments of novel microscale techniques such as nano-UPLC and improvements of mass spectrometry, the 3D separation becomes a reliable and practical selection. This review summarizes existing offline and online 3D-LC platforms developed for proteomics and their applications. In detail, setups and implementation of those systems as well as their advances are outlined. The performance of those platforms is also discussed and compared with the state-of-the-art 2D-LC. In addition, we provide some perspectives on the future developments and applications of 3D-LC in proteomics.
蛋白质组学是一种大规模的蛋白质研究，旨在描述和鉴定生物系统中所有表达的蛋白质。这些表达的蛋白质通常非常复杂，丰度范围也很大。为了实现蛋白质组分析的高准确性和灵敏度，多维（MD）分离和质谱的混合平台提供了最强大的解决方案。多维分离提供了更高的峰容量并减少了样品复杂性，使质谱能够以更高的灵敏度分析更多的蛋白质。尽管二维（2D）分离自蛋白质组学早期就已经广泛应用，但由于分离的重现性低、质谱分析时间增加，三维（3D）分离几乎没有被使用。随着新型微尺度技术（如纳升超高效液相色谱（nano-UPLC））的发展和质谱的改进，3D 分离成为一种可靠且实用的选择。本文综述了为蛋白质组学开发的现有离线和在线 3D-LC 平台及其应用。详细介绍了这些系统的设置和实施及其进展。还讨论了这些平台的性能，并与最先进的 2D-LC 进行了比较。此外，我们还对 3D-LC 在蛋白质组学中的未来发展和应用提供了一些看法。
