蛋白质/多肽的微流控二维芯片电泳*

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。     

基于毛细管电泳及微流控芯片的药物提取分离技术

近年来,在提取分离方面出现了许多新技术和新方法.其中毛细管电泳和微流控芯片技术以其微量、高效、快速等特点,在药物提取分离中已渐显优势.该文对基于毛细管电泳和微流控芯片的两相电泳技术、微流控液液萃取技术、微流控固液萃取技术、微流控过滤式分离技术、微流控膜分离技术在药物分离提取中的应用进行了综述.     

微流控芯片电泳的研究进展

该文综述了微流控芯片电泳的制备、结构和应用,比较了不同材料微流控芯片电泳的制备机理、表面改性和性能特点,归纳和总结了不同结构微流控芯片电泳的进样、分离和检测系统以及不同类型微流控芯片电泳在荧光物质、金属离子、糖、药物、核酸、DNA、氨基酸、多肽和蛋白质分析中的应用,并对微流控芯片电泳的未来发展方向做了展望.     

单细胞分析的研究

单细胞分析是分析化学、生物学和医学之间渗透发展形成的跨学科前沿领域。近年来,毛细管电泳及微流控芯片用于单细胞分析已取得显著进展,特别表现在微流控芯片用于细胞的培养、分选、操纵、定位、分离及检测细胞的组分,实时监测细胞释放,及高通量阵列检测等方面。芯片的单元操作可根据需要灵活组合,显示出其独特的优点。本文重点介绍作者研究组的工作,并对近三年来国内外在毛细管电泳及芯片毛细管电泳用于单细胞分析的新进展进行评论。最后从毛细管电泳与微流控芯片、微流控芯片与细胞界面以及量子点用于探测活细胞等方面,展望了单细胞分析的发展前景。     

顺序注射-液芯波导H-通道芯片联用系统DNA片段的毛细管电泳分离激光诱导荧光检测

毛细管电泳微流控芯片分离-激光诱导荧光(LIF)检测DNA片段是近年来微流控分析系统中研究得较为成功的领域,该方向的研究成果极大地促进了微流控分析系统的发展.在相关的报道中,待分析样品和系统运行溶液仍然主要使用手工操作.     

毛细管电泳和微流控芯片技术在临床尿检中的应用

本文就毛细管电泳和微流控芯片技术在临床尿检中的应用,以及毛细管电泳和微流控芯片技术在尿样前处理、样品富集方面的进展进行了综述。主要介绍了临床尿液一般化学检查和特殊化学检查,着重对肾功能指标的生化检查进行了总结。根据目前的研究状况,对毛细管电泳和微流控芯片技术在临床检测上的应用前景和发展方向进行了展望。     

微流控芯片电泳电导检测分离分析尿蛋白

基于自行构建的微流控芯片电泳集成非接触式电导检测分析系统,建立了一种集进样、分离与检测为一体的微流控芯片电泳电导检测蛋白质的方法,并用于人白蛋白(HSA)和人转铁蛋白(TRF)两种尿蛋白的分离分析以及肾病综合症病人尿液中白蛋白的定量检测.考察并优化了缓冲液、分离电压、进样方式、进样时间等电泳分离的影响因素,在缓冲液为p...     

毛细管电泳柱及微流控芯片通道涂层的发展

综述了用于毛细管电泳柱和微流控芯片通道的涂层材料和涂层技术的发展状况,以及涂层对分离效果和分离结果重现性的影响。将涂层材料按照动态和静态分类,静态涂层又分别按照均聚物、共聚物、杂环类等进行讨论;综述了交联反应法、溶胶-凝胶法、辐照法、化学沉积法等涂层的制备方法。对毛细管电泳柱和微流控芯片通道的改良具有一定的参考价值。     

纳米粒子毛细管电泳/微流控芯片新技术及其在手性分离中的应用

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。     

芯片电泳推拉式压力进样方法

芯片电泳作为微流控分析系统的典型代表,广泛涉及材料、微加工方法、微液流控制、分离模式和检测方法等诸多方面.与传统分析系统一样,样品制备和引入也是微全分析系统实现样品到结果首先面临的问题.电进样一直是芯片电泳系统的主流进样方法.而传统毛细管电泳系统中与电进样同样经常使用的压力进样方法则很少用于芯片电泳系统.     

Field amplified sample stacking coupled with chip-based capillary electrophoresis using negative pressure sample injection technique

A multi-T microchip for integrated field amplified sample stacking (FASS) with CE separation to increase the chip-based capillary electrophoresis (chip-based CE) sensitivity was developed. Volumetrically defined large sample plug was formed in one step within 5s by the negative pressure in headspace of the two sealed sample waste reservoirs produced using a syringe pump equipped with a 3-way valve. Stacking and separation can proceed only by switching the 3-way valve to release the vacuum in headspace of the two sample waste reservoirs. This approach considerably simplified the operations and the equipments for FASS in chip-based CE systems. Migration time precisions of 3.3% and 1.3% RSD for rhodamine123 (Rh123) and fluorescien sodium salt (Flu) in the separation of a mixture of Flu and Rh123 were obtained for nine consecutive determinations with peak height precisions of 4.8% and 3.4% RSD, respectively. Compared with the chip-based CE on the cross microchip, the sensitivity for analysis of FlTC, FITC-labeled valine (Val) and Alanine (Ala) increased 55-, 41- and 43-fold, respectively.     

Nanomaterials and chip-based nanostructures for capillary electrophoretic separations of DNA

Capillary electrophoresis (CE) and microchip capillary electrophoresis (MCE) using polymer solutions are two of the most powerful techniques for the analysis of DNA. Problems, such as the difficulty of filling polymer solution to small separation channels, recovering DNA, and narrow separation size ranges, have put a pressure on developing new techniques for DNA analysis. In this review, we deal with DNA separation using chip-based nanostructures and nanomaterials in CE and MCE. On the basis of the dependence of the mobility of DNA molecules on the size and shape of nanostructures, several unique chip-based devices have been developed for the separation of DNA, particularly for long DNA molecules. Unlike conventional CE and MCE methods, sieving matrices are not required when using nanostructures. Filling extremely low-viscosity nanomaterials in the presence and absence of polymer solutions to small separation channels is an alternative for the separations of DNA from several base pairs (bp) to tens kbp. The advantages and shortages of the use of nanostructured devices and nanomaterials for DNA separation are carefully addressed with respect to speed, resolution, reproducibility, costs, and operation.     

Analysis of inflammatory biomarkers from tissue biopsies by chip-based immunoaffinity CE

To aid in the biochemical analysis of human skin biopsies, a semiautomatic chip-based CE system has been developed for measuring inflammatory biomarkers in microdissected areas of the biopsy. Following solubilization of the dissected tissue, the desired biomarkers were isolated by immunoaffinity capture using a panel of 12 antibodies, immobilized on a disposable glass fiber disk, within the extraction port of the chip. The captured analytes were labeled with a 635 nm light-emitting laser dye and electroeluted into the separation channel. Electrophoretic separation of all of the analytes was achieved in 2.2 min with quantification of each peak being performed by online LIF detection and integration of each peak area. Comparison of the results obtained from the chip-based system to those obtained using commercially available high-sensitivity immunoassays demonstrated that the chip-based assay provides a fast, accurate procedure for studying the concentrations of inflammatory biomarkers in complex biological materials. The degree of accuracy and precision achieved by the chip-based CE is comparable to conventional immunoassays and the system is capable of analyzing circa six samples per hour. With the ever-expanding array of antibodies that are commercially available, this chip-based system can be applied to a wide variety of different biomedical analyses.     

Recent advances in CE and CEC of pollutants

Recent advances in CE and CEC separation, detection and sample preparation/preconcentration methodologies, for the determination of a variety of compounds having current or potential environmental relevance, have been overviewed. The reviewed literature has illustrated the wide range of CE applications available, indicating a continuing interest in CE and CEC in the environmental field. New developments in chip-based CE systems are also discussed.     

微流控芯片测定单细胞内化学组分的进展

细胞是生命的基本单元。由于细胞的个体差异,传统分析群体细胞的方法难以得到单细胞的重要信息。准确可靠地测定单细胞内化学组分的含量能大大提高从正常细胞中辨别不正常细胞的能力,为进一步研究和发展生物化学、医学和临床检验等领域奠定基础。近年来,用微流控芯片进行单细胞分析已引起广泛的兴趣。微流控芯片可以集成单细胞进样、溶膜、电泳分离胞内化学组分和高灵敏度测定等一系列操作步骤,为分析单细胞内的化学组分提供了新的技术平台。本文主要综述了近年来微流控芯片测定单细胞内化学组分的进展。重点在于利用电渗流、压力结合电渗流和激光镊子等技术操控单细胞在微流控芯片上完成单细胞进样、溶膜、细胞内化学组分的电泳分离和高灵敏度测定等一系列操作步骤。对在微流控芯片上的衍生技术也做了较为详细的阐述。     

Capillary electrophoresis-mass spectrometry for the analysis of intact proteins 2007-2010

CE coupled to MS has proven to be a powerful analytical tool for the characterization of intact proteins, as it combines the high separation efficiency of CE with the selectivity of MS. This review provides an overview of the development and application of CE-MS methods within the field of intact protein analysis as published between January 2007 and June 2010. Ongoing technological developments with respect to CE-MS interfacing, capillary coatings for CE-MS, coupling of CIEF with MS and chip-based CE-MS are treated. Furthermore, CE-MS of intact proteins involving ESI, MALDI and ICP ionization is outlined and overviews of the use of the various CE-MS methods are provided by tables. Representative examples illustrate the applicability of CE-MS for the characterization of proteins, including glycoproteins, biopharmaceuticals, protein-ligand complexes, biomarkers and dietary proteins. It is concluded that CE-MS is a valuable technique with high potential for intact protein analysis, providing useful information on protein identity and purity, including modifications and degradation products.     

Rapid fingerprinting of a highly glycosylated fusion protein by microfluidic chip-based capillary electrophoresis–mass spectrometry

Protein glycosylation can impact the efficacy, safety, and pharmacokinetics of therapeutic proteins. Achieving uniform and consistent protein glycosylation is an important requirement for product quality control at all stages of therapeutic protein drug discovery and development. The development of a new microfluidic CE device compatible with MS offers a fast and sensitive orthogonal mode of high-resolution separation with MS characterization. Here, we describe a fast and robust chip-based CE-MS method for intact glycosylation fingerprinting of a therapeutic fusion protein with complex sialylated N and O-linked glycoforms. The method effectively separates multiple sialylated glycoforms and offers a rapid detection of changes in glycosylation profile in 6 min.     

Rapid analysis of inflammatory cytokines in cerebrospinal fluid using chip-based immunoaffinity electrophoresis

A chip-based capillary electrophoresis system has been designed for rapidly measuring the concentrations of inflammatory cytokines in the cerebrospinal fluid of patients with head trauma. Isolation of the reactive cytokines was achieved by immunoaffinity capture using a panel of six immobilized antibodies, directly attached to the injection port of the chip. The captured cytokines were labeled in situ with a red light-emitting laser dye and electroeluted into the separation channel. Separation of the isolated cytokines was achieved by electrophoresis in under 2 min with quantification of the resolved peaks being achieved by on-line laser-induced fluorescence and integration of each peak area. Comparison of the results to commercially available high-sensitivity immunoassays demonstrates that the chip-based assay provides a fast, accurate procedure for studying the concentrations of these analytes in complex biological materials. The degree of accuracy and precision achieved by the chip-based CE is comparable to conventional immunoassays, the system being able to analyze between 10-12 samples per hour. With the ever-expanding array of antibodies that are commercially available, this chip-based system can be applied to a wide variety of different analyses.     

An automated electrokinetic continuous sample introduction system for microfluidic chip-based capillary electrophoresis

An automated and continuous sample introduction system for microfluidic chip-based capillary electrophoresis (CE) was developed in this work. An efficient world-to-chip interface for chip-based CE separation was produced by horizontally connecting a Z-shaped fused silica capillary sampling probe to the sample loading channel of a crossed-channel chip. The sample presentation system was composed of an array of bottom-slotted sample vials filled alternately with samples and working electrolyte, horizontally positioned on a programmable linearly moving platform. On moving the array from one vial to the next, and scanning the probe, which was fixed with a platinum electrode on its tip, through the slots of the vials, a series of samples, each followed by a flow of working electrolyte was continuously introduced electrokinetically from the off-chip vials into the sample loading channel of the chip. The performance of the system was demonstrated in the separation and determination of FITC-labeled arginine and phenylalanine with LIF detection, by continuously introducing a train of different samples. Employing 4.5 kV sampling voltage (1000 V cm(-1) field strength) for 30 s and 1.8 kV separation voltage (400 V cm(-1) field strength) for 70 s, throughputs of 36 h(-1) were achieved with <1.0% carryover and 4.6, 3.2 and 4.0% RSD for arginine, FITC and phenylalanine, respectively (n = 11). Net sample consumption was only 240 nL for each sample.     

Capacitively coupled contactless conductivity detection in capillary electrophoresis

Capacitively coupled contactless conductivity detection (C(4)D) has become an accepted detection method in capillary electrophoresis (CE) for a variety of analytes. Advantages of this technique over optical detection modes and galvanic contact conductivity detection include great flexibility in capillary handling and rather simple mechanical parts and electronics, as it can be performed in an on-capillary mode. Furthermore, the detection principle can be used with capillaries made of other materials than fused silica (PEEK, Teflon), with chip-based separation technologies, or with capillaries having very small inner diameters. This review presents a discussion of the published literature on C(4)D for CE and capillary electrochromatography.