采用多面体低聚倍半硅氧烷基杂化整体毛细管柱分离完整蛋白质。
Separation of intact proteins by using polyhedral oligomeric silsesquioxane based hybrid monolithic capillary columns.
机构信息
Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China; University of Chinese Academy of Sciences, Beijing 100049, China.
出版信息
J Chromatogr A. 2013 Nov 22;1317:138-47. doi: 10.1016/j.chroma.2013.09.004. Epub 2013 Sep 7.
High-efficient separation of intact proteins is still a huge challenge in proteome analysis of complex biological samples by using capillary columns. In this study, four POSS-based hybrid monolithic capillary columns were prepared and applied in nano-flow liquid chromatography (Nano-LC) separation of intact proteins. It was observed that the POSS-based hybrid monolithic columns exhibit high permeability, good LC separation reproducibility and column efficiency for intact protein separation. The effects of different LC separation conditions such as flow rate, gradient steepness, column length and mobile phase additives on the LC separation efficiency of the POSS-based hybrid monolithic column were systematically examined. Finally, fast LC separation of 7 proteins mixture was realized in 2.5 min by using the optimized conditions on the 100 μm i.d. POSS-based hybrid monolithic capillary column.
高效分离完整蛋白质仍然是通过毛细管柱对复杂生物样品进行蛋白质组分析的巨大挑战。本研究制备了四种基于 POSS 的杂化整体毛细管柱，并将其应用于完整蛋白质的纳流液相色谱（Nano-LC）分离。结果表明，基于 POSS 的杂化整体柱具有较高的渗透性、良好的 LC 分离重现性和完整蛋白质分离的柱效。系统考察了不同 LC 分离条件，如流速、梯度陡度、柱长和流动相添加剂对基于 POSS 的杂化整体柱 LC 分离效率的影响。最后，在 100μm i.d.基于 POSS 的杂化整体毛细管柱上优化条件下，实现了 7 种蛋白质混合物的快速 LC 分离，仅需 2.5 分钟。
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