共查询到18条相似文献,搜索用时 250 毫秒
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毛细管区带电泳中蛋白质吸附及其解决方法 总被引:1,自引:0,他引:1
本文评述毛细管区带电脉中蛋白质的吸附及解决办法,内容包括吸附的原因,通过缓冲溶液组成的变化、通过毛细管内壁改性、通过附加电场等方法降低吸附。对毛细管内壁电荷密度等方法降低吸附。尤其对毛细管内壁改性方法,改性对电渗流及分离效率的影响,改性的稳定性和重复性等进行了较为详细的阐述。 相似文献
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手性是自然界的本质属性之一。手性分离分析技术对生命科学、环境科学、生物工程和药物工程等许多学科都具有十分重要的意义。当前,对不同种类手性化合物进行拆分已成为毛细管电泳技术最具特色的研究和应用领域之一。然而,被分析物(或拆分剂)在毛细管内壁的吸附是毛细管电泳手性分离中的常见问题。涂层技术就是采用不同的方法对毛细管内壁进行改性,是抑制非特异性吸附、提高分离效率及分离重现性最简便和最有效的方法。本文主要综述了近十几年来各种涂层技术在毛细管电泳手性分离领域的应用现状,并对毛细管涂层技术今后的发展进行了展望。 相似文献
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用石英玻璃微珠模拟了HP-1,HP-20M,HP-17毛细管色谱柱内壁,用真空重量法测定了丙烷、丁烷、丙烯、液化石油气样品在3种模拟毛细管内壁表面的吸附等温线,并计算了等量吸附热随吸附量的变化关系。结果表明改性石英玻璃表面固定相的化学性质是吸附等温线类型的决定因素。本文所得结果对毛细管色谱动力学过程的理解,样品在毛细管柱上的活度系数及液化石油气组成准确测定等方面有重要意义。 相似文献
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聚合物涂层技术在毛细管电泳分离蛋白质中的应用 总被引:1,自引:0,他引:1
影响毛细管电泳分离蛋白质效率和重现性的主要原因是毛细管壁对蛋白质的吸附.本文综述了解决这一问题的途径,包括极端pH值法、添加小分子添加剂、对毛细管内壁改性等方法,重点介绍并比较了用于毛细管内壁改性的两类聚合物涂层-化学键合和物理吸附的涂层.分别讨论了两类涂层在对电渗流的抑制和调节、对蛋白质吸附的阻止等方面的作用效果,并且对同种涂覆机理下不同聚合物的涂覆效果做了相关比较.总体上说,物理吸附的涂层比化学键合的涂层性能优越,因此在毛细管涂覆领域更受欢迎.尽管目前该领域的工作取得了较大进展,但仍未找到一种对所有种类蛋白质的分离都有效的普适性涂层,因此未来的一段时间内分离复杂组分的蛋白质仍面临着挑战. 相似文献
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毛细管电泳具有分析时间短,分离效率高,样品消耗量少等优点,在生物样品分离,特别是蛋白质分析领域有重要应用。然而,毛细管内壁硅羟基的解离给分离结果带来诸多不良影响。聚合物涂层能够抑制蛋白质在毛细管内壁的吸附以及调控电渗流,故对毛细管内壁进行有效修饰能够提高其对蛋白质的分离效率及分离稳定性。该文主要综述了动态及静态聚合物涂层毛细管的最新研究进展,并概述了近些年基于多巴胺/聚多巴胺发展起来的涂层毛细管的研究进展,最后展望了聚合物涂层毛细管的发展趋势。 相似文献
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碱性蛋白质毛细管电泳分离研究 总被引:1,自引:0,他引:1
碱性蛋白质毛细管电泳分离研究任吉存,邓延倬,程介克(武汉大学化学系分析测试科学系,武汉,430072)关键词毛细管电泳,碱性蛋白质,吸附,精氨酸,赖氨酸蛋白质的吸附作用严重影响了毛细管电泳分离蛋白质的重现性[1].为此,人们寻找各种途径来克服蛋白质的... 相似文献
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Capillary electrophoresis (CE) provides a new analytical tool for the separation of proteins, and almost all traditional modes of electrophoresis can be carry out in CE. But serious adsorption of proteins on capillary wall prohibited the proper separation. Three main approaches are used to overcome adsorption and control electroosmotic flow, (1) buffer of high or low pH,high salt concentration and additives, (2) pre-adsorption of neutral or charged macromolecules on the capillary wall and (3) chemically bonded coatings which are expected to give the best shield of silanol groups present on bare silica by vaious hydrophilic polymers. Capillaries coated with linear polyacrylamide represent the most successful approach available to date. Cross-linkage of polyacrylamide coating is desired to increase its stability. 相似文献
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径向电场调制毛细管电泳法用于蛋白质分离 总被引:1,自引:0,他引:1
利用自制的双向电场控制毛细管电泳新系统,考察了蛋白质的分离情况.结果发现,在低pH值下,通过施加径向电场,不仅可改变电渗流的大小和方向,而且能抑制蛋白质的吸附,进而实现对蛋白质分离效率和分离速度的调控.研究结果表明,可通过物理化学方法实现毛细管电泳的动态或随机调控,这对许多生物样品分离有实际意义. 相似文献
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δ‐Gluconolactone was covalently coupled with aminopropyl‐derivatized capillary, creating hydrophilic brushes on the inner wall of the capillary. The hydrophilic coating provided suppression of EOF and minimized protein adsorption, resulting in the separation of basic proteins and DNA with efficiencies up to 450 000 plates/m. The intra‐ and inter‐day repeatabilities of the coating referring to the migration times of the four tested proteins were satisfactory with RSD of no more than 1.1 and 1.8% (n=5), respectively. Two hundred consecutive runs were performed with negligible change in migration times and efficiency. 相似文献
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A simple coating technique by using uncross-linked dextran has been developed for fused-silica capillaries to be used in capillary electrophoresis of basic proteins. The capillaries were first silanized with a heterobifunctional silane (gamma-aminopropyltriethoxylsilane), which served as a coupling agent between the capillary inner wall and the polysaccharide coating. Dextran of high molecular mass (about 70 kDa) was activated with 1,1'-carbonyldiimidazole. Then the activated dextran was coupled to the primary amino groups that were anchored onto the inner wall of the silanized capillaries. The residual reactive groups on the dextran were further substituted by neutral functions in a coupling reaction with excess ethanolamine. By using dimethyl sulfoxide (DMSO) rather than aqueous buffer as the reaction medium, the extent of substitution was improved by minimizing the residual reactive groups at the surface. Since they are ionogenic, the electrosmotic flow in the system is relatively low. The chemically bound dextran coating showed good reproducibility and stability. In electrophoretic experiments basic proteins were separated with high efficiency by use of the dextran-coated fused-silica capillary columns. The main advantage of the method described here is that both polysaccharide activation and amine-coupling reactions were carried out under mild conditions at room temperature without catalysts. For this reason, the method is recommended to coat the inner wall of microfluidic separation channels which would not tolerate a harsh treatment. 相似文献
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Surface modification of the inner capillary wall in CE of proteins is frequently required to alter EOF and to prevent protein adsorption. Manual protocols for such coating techniques are cumbersome. In this paper, an automated covalent linear polyacrylamide coating and regeneration process is described to support long‐term stability of fused‐silica capillaries for protein analysis. The stability of the resulting capillary coatings was evaluated by a large number of separations using a three‐protein test mixture in pH 6 and 3 buffer systems. The results were compared to that obtained with the use of bare fused‐silica capillaries. If necessary, the fully automated capillary coating process was easily applied to regenerate the capillary to extend its useful life‐time. 相似文献