Anal. Chem.：强阳离子交换整体柱在蛋白质组学分析中应用新进展

佚名

美国化学会刊物Journal of Proteome Research 10月(2007，6，3872)的Currents栏目发表了题为“Monolithic SCX column for shotgun proteomics” 的新闻报道。该报道介绍了中科院大连化物所王方军、董靖、叶明亮和邹汉法研究员等人关于强阳离子交换整体柱在蛋白质组学中的应用方面的研究进展。作为蛋白质组学研究领域的权威刊物，JPR每期从各种重要的国际性学术刊物中挑选5篇左右的关于蛋白质组学方面的研究论文作为重要新闻在Currents栏目中进行报道。

国际上普遍采用带磺酸基团的单体合成强阳离子交换整体柱进行离子色谱分离分析，而这种类型的整体柱在不同分离缓冲溶液中容易发生溶胀或收缩，且在分离过程中可能掺杂有较强的疏水相互作用，因此不利于在多维色谱分离方面的应用。邹汉法研究员等人首次采用带磷酸基团的单体制备成功了一种新型强阳离子交换整体柱材料，该整体柱材料具有性质稳定、柱压降低、传质阻力小、离子交换容量高的特点，且表现出单一的阳离子交换机理。采用所制备的整体柱作为预柱，以两相柱进样和柱切换的原理建立新的接口模式，成功发展出高性能的用于纳升级RPLC-MS/MS分析的自动进样和柱切换多维分离技术。该系统有效减少进样时间、进样过程中的样品损失，大大提高了分离效率和分离的通量。该新型阳离子交换整体作为预柱被应用于在线多维色谱分离复杂的酵母蛋白酶解物，通过shotgun的技术，鉴定得到了1522个蛋白质(假阳性率仅为0.46%)。该研究成果9月份发表在美国化学会刊物Analytical Chemistry (AC)上(Anal. Chem. 2007, 79, 6599-6606)。（大连化物所）

原始出处:

Anal. Chem., 79 (17), 6599 -6606, 2007. 10.1021/ac070736f S0003-2700(07)00736-6
Web Release Date: August 4, 2007 Copyright © 2007 American Chemical Society

Capillary Trap Column with Strong Cation-Exchange Monolith for Automated Shotgun Proteome Analysis

Fangjun Wang, Jing Dong, Xiaogang Jiang, Mingliang Ye, and Hanfa Zou*

National Chromatographic Research and Analysis Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China

Received for review April 13, 2007. Accepted June 26, 2007.

Abstract:

A 150 m internal diameter capillary monolithic column with a strong cation-exchange stationary phase was prepared by direct in situ polymerization of ethylene glycol methacrylate phosphate and bisacrylamide in a trinary porogenic solvent consisting dimethylsulfoxide, dodecanol, and N,N'-dimethylformamide. This phosphate monolithic column exhibits higher dynamic binding capacity, faster kinetic adsorption of peptides, and more than 10 times higher permeability than the column packed with commercially available strong cation-exchange particles. It was applied as a trap column in a nanoflow liquid chromatography-tandem mass spectrometry system for automated sample injection and online multidimensional separation. It was observed that the sample could be loaded at a flow rate as high as 40 L/min with a back pressure of ~1300 psi and without compromising the separation efficiency. Because of its good orthogonality to the reversed phase separation mechanism, the phosphate monolithic trap column was coupled with a reversed-phase column for online multidimensional separation of 19 g of the tryptic digest of yeast proteins. A total of 1522 distinct proteins were identified from 5608 unique peptides (total of 54 780 peptides) at the false positive rate only 0.46%.