High-resolution separations of protein isoforms with liquid chromatography time-of-flight mass spectrometry using polymer monolithic capillary columns
High-resolution separations of protein isoforms with liquid chromatography time-of-flight mass spectrometry using polymer monolithic capillary columns
复制标题
DOI：
10.1016/j.chroma.2011.06.049
复制
发表时间：
2011-08-12
影响因子：
4.1
通讯作者：
Treumann, Achim
中科院分区：
文献类型：
--
作者：
Eeltink, Sebastiaan;Wouters, Bert;Treumann, Achim
完整的蛋白质，包括蛋白质异构体所产生的各种氨基酸的修改，采用聚（苯乙烯-co-二乙烯基苯）的整体毛细管柱在高效液相色谱耦合在线的飞行时间质谱仪（MS）的分离。使用250 mm x 0.2 mm整体毛细管柱，在2小时线性梯度下实现了峰容量>600的高灵敏度分离，并在移动的相中加入甲酸作为离子配对剂。高分辨率色谱与高准确度MS的组合允许区分仅在其氧化和生物素化状态方面不同的蛋白质同种型，从而允许分离结构同种型。最后，潜在的分离蛋白质异构体由于糖基化进行了讨论。(C)2011 Elsevier B. V.保留所有权利。
The separation of intact proteins, including protein isoforms arising from various amino-acid modifications, employing a poly(styrene-co-divinylbenzene) monolithic capillary column in high-performance liquid chromatography coupled on-line to a time-of-flight mass spectrometer (MS) is described. Using a 250 mm x 0.2 mm monolithic capillary column high-sensitivity separations yielding peak capacities of >600 were achieved with a 2 h linear gradient and formic acid added in the mobile phase as ion-pairing agent. The combination of high-resolution chromatography with high-accuracy MS allowed to distinguish protein isoforms that differ only in their oxidation and biotinylation state allowing the separation between structural isoforms. Finally, the potential to separate proteins isoforms due to glycosylation is discussed. (C) 2011 Elsevier B.V. All rights reserved.