美國化學(xué)會刊物Journal of Proteome Research 10月(2007,，6,，3872)的Currents欄目發(fā)表了題為“Monolithic SCX column for shotgun proteomics” 的新聞報道,。該報道介紹了中科院大連化物所王方軍、董靖,、葉明亮和鄒漢法研究員等人關(guān)于強(qiáng)陽離子交換整體柱在蛋白質(zhì)組學(xué)中的應(yīng)用方面的研究進(jìn)展,。作為蛋白質(zhì)組學(xué)研究領(lǐng)域的權(quán)威刊物，JPR每期從各種重要的國際性學(xué)術(shù)刊物中挑選5篇左右的關(guān)于蛋白質(zhì)組學(xué)方面的研究論文作為重要新聞在Currents欄目中進(jìn)行報道,。
國際上普遍采用帶磺酸基團(tuán)的單體合成強(qiáng)陽離子交換整體柱進(jìn)行離子色譜分離分析,，而這種類型的整體柱在不同分離緩沖溶液中容易發(fā)生溶脹或收縮，且在分離過程中可能摻雜有較強(qiáng)的疏水相互作用,，因此不利于在多維色譜分離方面的應(yīng)用,。鄒漢法研究員等人首次采用帶磷酸基團(tuán)的單體制備成功了一種新型強(qiáng)陽離子交換整體柱材料，該整體柱材料具有性質(zhì)穩(wěn)定,、柱壓降低,、傳質(zhì)阻力小、離子交換容量高的特點,，且表現(xiàn)出單一的陽離子交換機(jī)理,。采用所制備的整體柱作為預(yù)柱，以兩相柱進(jìn)樣和柱切換的原理建立新的接口模式,，成功發(fā)展出高性能的用于納升級RPLC-MS/MS分析的自動進(jìn)樣和柱切換多維分離技術(shù),。該系統(tǒng)有效減少進(jìn)樣時間,、進(jìn)樣過程中的樣品損失，大大提高了分離效率和分離的通量,。該新型陽離子交換整體作為預(yù)柱被應(yīng)用于在線多維色譜分離復(fù)雜的酵母蛋白酶解物,，通過shotgun的技術(shù)，鑒定得到了1522個蛋白質(zhì)(假陽性率僅為0.46%),。該研究成果9月份發(fā)表在美國化學(xué)會刊物Analytical Chemistry (AC)上(Anal. Chem. 2007, 79, 6599-6606),。（大連化物所）
原始出處:
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%.
