Rapid capillary electrophoresis time-of-flight mass spectrometry separations of peptides and proteins using a monoquaternarized piperazine compound (M7C4I) for capillary coatings
Rapid capillary electrophoresis time-of-flight mass spectrometry separations of peptides and proteins using a monoquaternarized piperazine compound (M7C4I) for capillary coatings
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DOI：
10.1002/elps.200700737
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发表时间：
2008-04-01
期刊：
影响因子：
2.9
通讯作者：
Bergquist, Jonas
中科院分区：
文献类型：
--
作者：
Elhamili, Anisa;Wetterhall, Magnus;Bergquist, Jonas
单季铵化哌嗪，1-（4-碘丁基）4-氮杂-1-氮鎓双环[2，2，2]辛烷碘化物（M7 C4 I），已被评估为表面衍生化试剂，用于CE结合TOF MS分析蛋白质，肽，和蛋白质类药物。M7 C4 I哌嗪在碱性pH值下通过离子化硅烷醇的烷基化形成共价键，产生具有高度稳定和反向极性的阳离子表面。所获得的表面在酸性pH下产生肽和蛋白质的快速分离（小于5分钟），具有高分离效率（对于肽高达1.1 × 10（6）板/m，对于蛋白质高达1.8 × 10（6）板/m），并且没有观察到涂层试剂渗入质谱仪。如果需要，涂覆程序的简单性还使得表面能够快速（2分钟）再生。这在复杂样品的分析中很有用，以防止可能的记忆效应。通过分离肽、蛋白质和蛋白质底物，证明了使用M7 C4 I涂层毛细管对复杂样品进行MS分析的潜力。更重要的是，可以分离分子量超过0.5 MDa的大型完整蛋白质，这是一件壮观的事情。如IgG（1）（150 kDa）和甲状腺球蛋白（669 kDa）的分离所证明的，涂层显示出以高效率处理这些大蛋白质的良好能力并保留峰形。
A monoquaternarized piperazine, 1-(4-iodobutyl) 4-aza-1-azoniabicyclo[2,2,2] octane iodide (M7C4I), has been evaluated as a surface derivatization reagent for CE in combination with TOF MS for the analysis of proteins, peptides, and protein digests. The M7C4I piperazine, at alkaline pH, forms a covalent bond via alkylation of the ionized silanols producing a cationic surface with a highly stable and reversed EOF. The obtained surface yields rapid separations (less than 5 min) of peptides and proteins at acidic pH with high separation efficiencies (up to 1.1 X 10(6) plates/m for peptides and up to 1.8 x 10(6) plates/m for proteins) and no observed bleeding of the coating reagent into the mass spectrometer. The simplicity of the coating procedure also enables fast (2 min) regeneration of the surface, if necessary. This is useful in the analysis of complex samples in order to prevent possible memory effects. The potential of using M7C4I-coated capillaries for MS analysis of complex samples is demonstrated by the separation of peptides, proteins, and protein digests. Even more, the spectacular thing in which large intact proteins with molecular masses over 0.5 MDa could be separated. The coating showed good ability to handle these large proteins with high efficiency and retained peak shape as demonstrated by separation of IgG(1) (150 kDa) and thyroglobulin (669 kDa).