CAS Key Lab of Separation Sciences for Analytical Chemistry, National Chromatographic Research and Analysis Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China.
Methods Mol Biol. 2022;2500:43-53. doi: 10.1007/978-1-0716-2325-1_5.
High-performance separation of proteoforms plays an important role in top-down proteomic ananlysis due to high complexity of the proteome. To this end, the functionalized ethylene-bridged hybrid monolithic materials have been developed for reversed-phase liquid chromatographic separation of proteoforms followed by online combination with high-resolution mass spectrometry (MS) for top-down proteomic analysis. Such monoliths have advantages of homogenously distributed functional groups in the framework, good chemical stability, and high permeability and, thus, show high resolution, good reproducibility, and low backpressure for proteoform separation. This chapter describes in detail the preparation of such monoliths and online combination with high-resolution MS for proteoform separation and identification.
由于蛋白质组的高度复杂性，在从头蛋白质组分析中，蛋白质形式的高效分离起着重要作用。为此，开发了功能化的乙烯桥联杂化整体材料，用于反相液相色谱分离蛋白质形式，然后与高分辨率质谱（MS）在线组合，用于从头蛋白质组分析。这种整体材料具有在骨架中均匀分布的功能基团、良好的化学稳定性、高渗透性等优点，因此对蛋白质形式的分离具有高分辨率、良好的重现性和低背压。本章详细描述了这种整体材料的制备以及与高分辨率 MS 的在线组合，用于蛋白质形式的分离和鉴定。
