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微流控芯片毛细管电泳在蛋白质分离分析中的应用研究进展 总被引:1,自引:1,他引:0
重点介绍了近年来国内外在微流控芯片毛细管电泳法用于蛋白质分离分析方面的研究进展。按照分离模式的不同,综述了各种应用于蛋白质分离的微流控芯片毛细管电泳系统,讨论了抑制芯片中的蛋白吸附的各种方法,并展望了芯片毛细管电泳系统在蛋白质分离领域的发展前景。引用文献47篇。 相似文献
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纳米粒子因其具有较大的比表面积和良好的生物相容性等特点,已广泛应用于分离科学领域。纳米粒子毛细管电泳/微流控芯片技术是纳米材料技术与毛细管电泳/微流控芯片技术相结合的产物。纳米粒子可以被吸附或键合到毛细管壁作为固定相与被分析物发生相互作用;也可以作为假固定相参与样品在柱内的分配和保留,从而提高柱效,改善分离。手性是自然界的本质属性之一,开发新的快速、高效、灵敏的手性分离分析方法对于对映体的立体选择性合成、药理研究、手性纯度检测和环境检测都具有重要的意义。本文主要综述了近些年来几种不同类型纳米粒子(聚合物纳米粒子、磁性纳米粒子、金纳米粒子、碳纳米管和其他类型纳米粒子)用于毛细管电泳/微流控芯片进行手性分离的现状,并对该领域今后的发展进行了展望。 相似文献
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在微流控芯片上构建多维分离系统,为蛋白质组学研究提供了一个有发展前景的高效分离分析技术平台。本文介绍了二维芯片电泳系统耦联模式选取及正交性评价的方法;综述了针对蛋白质/多肽分离分析的各种耦联模式微流控二维芯片电泳分析系统,如胶束电动力学色谱(MEKC)与毛细管区带电泳(CZE),开管电色谱(OECE)与CZE,等电聚焦(IEF)与CZE, IEF与SDS毛细管凝胶电泳(CGE), SDS-CGE与MEKC等。特别对二维电泳芯片切换接口的类型进行了分类,探讨了用于微流控二维芯片电泳系统的检测技术,并展望了微流控二维电泳芯片在蛋白质组学研究中的应用前景和发展方向。 相似文献
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毛细管电泳(CE)具有分离时间短、分离效率高、样品消耗量低等优点,在分离分析领域有着重要应用。原始的未修饰熔融石英毛细管只能提供阴极流向的电渗流和单一的电泳分离机制,分离性能有限,重复性较差,不能满足各类复杂样品体系尤其是中性和手性样品的分离需求。因此,有必要在CE中引入各类毛细管修饰策略,以拓展其实际应用潜力。贻贝仿生聚多巴胺(PDA)及其衍生材料因其简便易行的制备过程、优异的表面黏附性、良好的生物相容性、较强的二次反应活性和化学稳定性等优点,在催化、传感、水处理、样品前处理、生物医药以及CE分离等领域得到了广泛应用。PDA涂层的制备过程与物理吸附涂层一样简便,而表面黏附涂层的稳定性又可与共价键合涂层相媲美,因此非常适用于石英毛细管柱的修饰。更重要的是,PDA涂层较强的二次反应活性使其可作为反应平台进行灵活多样的二次表面修饰,便于构建多功能PDA涂层毛细管电色谱(CEC)固定相。基于这些突出优点,PDA涂层材料在CEC中的巨大应用价值逐渐得到了研究者们的广泛关注。该文首先对近3年有关PDA形成机理及PDA快速沉积表面化学的最新研究进展进行了总结,在此基础上综述了近10年PDA涂层材料在开管毛细管电色谱(OT-CEC)和毛细管电色谱整体柱中的最新应用。此外,还对PDA涂层材料在CEC中的发展方向进行了展望。 相似文献
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A novel method for the preparation of covalently linked capillary coatings of PVA was demonstrated using photosensitive diazoresin (DR) as coupling agents. Layer‐by‐layer self‐assembly film of DR and PVA based on hydrogen bonding was first fabricated on the inner wall of capillary, then the hydrogen bonding was converted into covalent bonding after treatment with UV light through the unique photochemistry reaction of DR. The covalently bonded coatings suppressed basic protein adsorption on the inner surface of capillary, and thus a baseline separation of lysozyme, cytochrome c and BSA was achieved using CE. Compared with bare capillary or noncovalently bonded DR/PVA coatings, the covalently linked DR/PVA capillary coatings not only improved the CE separation performance for proteins, but also exhibited good stability and repeatability. Due to the replacement of highly toxic and moisture‐sensitive silane coupling agent by DR in the covalent coating preparation, this method may provide a green and easy way to make the covalently coated capillaries for CE. 相似文献
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Mussel‐inspired polydopamine has been widely used in capillary electrophoresis as a facile and universal tool for the surface modification of capillary, mainly due to its versatility, stability, strong adhesiveness, and biocompatibility properties. In this review, the recent development of mussel‐inspired surface chemistry with rapid deposition of polydopamine was introduced, and the recent applications of polydopamine in capillary electrophoresis (2011–July 2018) were summarized into four main aspects, namely, sample pretreatments, functional coatings, codeposition coatings, and intermediate coatings. Further study may be focused on clarifying the mechanisms of polydopamine formation and polydopamine‐assisted codeposition, and expanding coatings methods to plant polyphenols. 相似文献
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《Journal of separation science》2017,40(20):4060-4066
Capillary electrophoresis and electrokinetic chromatography are typically carried out in unmodified fused‐silica capillaries under conditions that result in a strong negative zeta potential at the capillary wall and a robust cathodic electroosmotic flow. Modification of the capillary wall to reverse the zeta potential and mask silanol sites can improve separation performance by reducing or eliminating analyte adsorption, and is essential when conducting electrokinetic chromatography separations with cationic latex nanoparticle pseudo‐stationary phases. Semipermanent modification of the capillary walls by coating with cationic polymers has proven to be facile and effective. In this study, poly([2‐(acryloyloxy)ethyl]trimethylammonium chloride) polymers were synthesized by reversible addition‐fragmentation chain transfer polymerization and used as physically adsorbed semipermanent coatings for capillary electrophoresis and electrokinetic chromatography separations. An initial synthesis of poly([2‐(acryloyloxy)ethyl]trimethylammonium chloride) polymer coating produced strong and stable anodic electroosmotic flow of –5.7 to –5.4 × 10−4 cm2/V⋅s over the pH range of 4–7. Significant differences in the magnitude of the electroosmotic flow and effectiveness were observed between synthetic batches, however. For electrokinetic chromatography separations, the best performing batches of poly([2‐(acryloyloxy)ethyl]trimethylammonium chloride) polymer performed as well as the commercially available cationic polymer polyethyleneimine, whereas polydiallylammonium chloride and hexadimethrine bromide did not perform well. 相似文献
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Protein adsorption to the inner capillary wall hinders the use of kinetic capillary electrophoresis (KCE) when studying noncovalent protein-ligand interactions. Permanent and dynamic capillary coatings have been previously reported to alleviate much of the problems associated with protein adsorption. The characteristic limitations associated with permanent and dynamic coatings motivated us to look at a third type of coating - semipermanent. Here, we demonstrate that a semipermanent capillary coating, designed by Lucy and co-workers, comprised of dioctadecyldimethylammonium bromide (DODAB) and polyoxyethylene (POE) stearate, greatly reduces protein adsorption at physiological pH - a necessary requirement for KCE. The coating (i) does not inhibit protein-DNA complex formation, (ii) prevents the adsorption of the analytes, and (iii) supports an electoosmotic flow required for many applications of KCE. The coating was tested in three physiological buffers using a well-known DNA aptamer and four proteins that severely bind to bare silica capillaries as standards. For every protein, a condition was found under which the semipermanent coating effectively suppresses protein adhesion. While no coating can completely prevent the adsorption of all proteins, our findings suggest that the DODAB/POE stearate coating can have a broad impact on CE at large, as it prevents the absorption of several well studied, highly adhesive proteins at physiological pH. 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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Polyelectrolytes are widely used in capillary electrophoresis as coating agents of silica capillaries to prevent adsorption phenomena and improve the repeatability of peptide and protein analysis. A systematic study of the coating experimental conditions has been carried out to optimize coating stability and performance. The main experimental parameters studied were the type and concentration of polyelectrolytes used in several monolayer and multilayer coatings, the ionic strength of coating and stabilizing solutions, and the procedures used for coating and capillary storage. Electroosmotic flow magnitude, direction and repeatability were used to monitor coating stability. Coating ability to limit adsorption was investigated by monitoring variations of migration times, time-corrected peak areas and separation efficiency of test peptides. Capillary-to-capillary and batch-to-batch reproducibility was also studied. In addition, the separation performance of polyelectrolyte coatings were compared to those obtained with bare silica capillaries. 相似文献