Reversed-phase electrochromatography of amino acids and peptides using porous polymer monoliths.
Reversed-phase electrochromatography of amino acids and peptides using porous polymer monoliths.
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使用多孔聚合物整料对氨基酸和肽进行反相电色谱分析。
DOI：
10.1016/s0021-9673(01)01036-6
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发表时间：
2001
期刊：
影响因子：
--
通讯作者：
A. K. Singh
中科院分区：
文献类型：
--
作者：
R. Shediac;S. Ngola;D. Throckmorton;D. Anex;T. Shepodd;A. K. Singh
高效快速地分离微量氨基酸和生物活性肽在新兴的蛋白质组学领域以及临床和制药领域具有重要意义。我们开发了新型紫外线引发丙烯酸酯基多孔聚合物整体作为固定相，用于阳离子、阴离子和中性氨基酸和肽的毛细管和芯片电色谱，然后进行吸光度或激光诱导荧光检测。刚性整体是铸造成型的，并且可以调节电荷和疏水性。对于低pH值的分离，使用含有季胺部分的整体材料来实现高电渗流，对于高pH值的分离，使用具有酸性磺酸基团的整体材料。使用毛细管中带负电和带正电的聚合物整料，可以高效且可重复地分离苯硫基乙内酰脲标记的氨基酸、天然肽以及用萘-2,3-二甲醛 (NDA) 标记的氨基酸和肽。通过毛细管电色谱法获得的分离效率范围为 65 000–371 000 板/米。系统地研究了缓冲液组成和柱疏水性程度以优化分离。还将整料浇铸在玻璃芯片的微通道中，并进行电色谱分离，然后进行激光诱导荧光检测，获得了三种 NDA 标记的生物活性肽。
Efficient and rapid separation of minute levels of amino acids and bioactive peptides is of significant importance in the emerging field of proteomics as well as in the clinical and pharmaceutical arena. We have developed novel UV-initiated acrylate-based porous polymer monoliths as stationary phases for capillary- and chip-electrochromatography of cationic, anionic, and neutral amino acids and peptides, followed by absorbance or laser-induced fluorescence detection. The rigid monoliths are cast-to-shape and are tunable for charge and hydrophobicity. For separations at low pH, monoliths containing quaternary amine moieties were used to achieve high electroosmotic flow, and for high pH separations monoliths with acidic sulfonic acid groups were employed. Efficient and reproducible separations of phenylthiohydantoin-labeled amino acids, native peptides, and amino acids and peptides labeled with naphthalene-2,3-dicarboxaldehyde (NDA) were achieved using both negatively- and positively-charged polymer monoliths in capillaries. Separation efficiencies in the range of 65 000–371 000 plates/m were obtained with capillary electrochromatography. Buffer composition and the degree of column hydrophobicity were studied systematically to optimize separations. The monoliths were also cast in the microchannels of glass chips and electrochromatographic separation followed by laser-induced fluorescence detection of three NDA-labeled bioactive peptides was obtained.
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采用在线电喷雾电离和飞行时间质谱法对生物分子进行毛细管电色谱分析。
DOI：
10.1016/s0021-9673(98)00817-6
发表时间：
1998
期刊：
Journal of chromatography. A
影响因子：
--
作者：
Choudhary,G;Horváth,C;Banks,JF
通讯作者：
Banks,JF
影响因子：
7.4
作者：
Huber,CG;Choudhary,G;Horváth,C
通讯作者：
Horváth,C
影响因子：
7.4
作者：
Peters, EC;Petro, M;Fréchet, JMJ
通讯作者：
Fréchet, JMJ
使用阳离子丙烯酸整料对蛋白质和肽进行毛细管电色谱分析。
DOI：
10.1016/s0021-9673(00)00250-8
发表时间：
2000
期刊：
Journal of chromatography. A
影响因子：
--
作者：
Zhang,S;Huang,X;Zhang,J;Horváth,C
通讯作者：
Horváth,C
影响因子：
7.4
作者：
E. C. Peters;M. Petro;F. Švec;J. Fréchet
通讯作者：
E. C. Peters;M. Petro;F. Švec;J. Fréchet