毛细管等电聚焦电泳技术进展

介绍了毛细管等电聚焦电泳(CIEF)技术的发展和现状,特别对载体两性电解质、柱内表面处理和检测技术等方面的最新进展加以系统评述,并且对CIEF的定性能力、峰容量、重现性及其在生命科学方面的应用前景进行了讨论。引用文献59篇。     

毛细管等电聚焦和电渗泵驱动聚焦区带分离蛋白质

建立了一种利用电渗泵驱动毛细管内的聚焦区带,实现毛细管电泳等电聚焦分离蛋白质的方法。通过控制电压来调节泵的输出流量,从而调节聚焦区带的迁移速度。适用于毛细管电泳等电聚焦两步法分离蛋白质等两性物质。考察了对牛血清白蛋白和溶菌酶两种粗提蛋白质混合物的分离,迁移时间的RSD分别为1.6%和1.3%,峰面积的RSD均为1.6%,证明方法可行。     

毛细管等电聚焦分离蛋白质

采用均匀设计法进行未涂壁石英毛细管等电聚焦研究,考察两性电解质、甲基纤维素、四甲基乙二胺和牛磺酸几种组分对等电聚焦的影响,确定出了比较合适的浓度范围。当中性蛋白质迁移通过检测窗后,在进样端施加一低气压,从而使酸性蛋白质得到很好的分离,适用于较宽的ｐＩ范围。方法的重复性好,为蛋白质分离鉴定提供一种有力的工具。     

毛细管等电聚焦/加压毛细管电色谱二维分离体系在多肽分离中的应用

以毛细管等电聚焦(cIEF)为第一维分离模式,以反相加压毛细管电色谱(pCEC)为第二维分离模式,开展离线二维色谱分离研究,并对复杂肽段进行分离.羟丙基纤维紊(HPC)涂层的毛细管用于cIEF分离,对6种标准蛋白质的平均分离柱效约为31万.在毛细管末端引进电隔离槽,方便了第一维样品的收集.在加电6 kV下,第二维pCE...     

毛细管等电聚焦/毛细管无胶筛分电泳二维蛋白质分离平台的构建

设计并制作了一种新型的中空纤维接口,以此为核心构建了毛细管等电聚焦(CLEF)／毛细管无胶筛分(CNGE)电泳二维蛋白质分离技术平台,实现了将二维凝胶电泳从平板转移到毛细管中。利用该二维分离平台,对血红蛋白样品进行了高效、快速分离分析,验证了该二维分离平台的可行性及分离效能。实验结果表明：二维分离系统总的峰个数和分离效能都比各自一维分离系统有较大提高。     

毛细管等电聚焦一毛细管区带电泳二维蛋白质分离平台的构建及其性能

通常采用二维聚丙烯酰胺凝胶电泳(2D-PAGE)分析组织或细胞的全蛋白质，但难与质谱(MS)直接联用．用高效液相色谱(HPLC)和毛细管电泳(CE)分离分析蛋白质和多肽的一维分离模式的分辨率和峰容量有限．多维柱联用技术比一维分离有更高的分辨率和峰容量，便于和MS直接联用，     

新型固定化pH梯度毛细管等电聚焦方法用于蛋白分离

通过化学键合建立一种固定化pH梯度的方法,用于毛细管等电聚焦分离蛋白质.采用微流控泵驱动毛细管内的聚焦区带,通过调节泵的流量,从而调节聚焦区带的迁移速度.该方法避免了自由溶液聚焦时两性电解质所带来的影响,实现了高灵敏度及检测波长自由选择等优点,适用于两步法毛细管电泳等电聚焦分离蛋白质等两性电解质.本文考察了对牛血清白蛋白和血红蛋白两种蛋白质混合物的分离,证明了该方法可行.     

微等电聚焦分离等位基因表达蛋白的研究

借助于显微操作方法,用等电聚焦和改进的超敏感染色技术对西葫芦单个花粉粒中的可溶性蛋白进行了电泳分析,获得了清晰的电泳图谱。在靠近酸性端有两条相距极近而且分布比较特殊的蛋白带,在第１种植株中,这两条带稍微靠近酸性端;在第２种植株中,它们稍微远离酸性端;在第３种植株中,它们相同于第１种和第２种,而且在两种之间的分布比例为１∶１。就这两条带的分布情况在判断植株的相对纯合性或杂合性方面的应用以及单花粉蛋白等电聚焦技术配合显微操作方法在群体遗传学和发育生物学研究方面的应用作了讨论。     

毛细管等电聚焦-毛细管区带电泳二维蛋白质分离平台的构建及其性能

通常采用二维聚丙烯酰胺凝胶电泳 ( 2 D- PAGE)分析组织或细胞的全蛋白质 [1] ,但难与质谱 ( MS)直接联用 .用高效液相色谱 ( HPLC)和毛细管电泳 ( CE)分离分析蛋白质和多肽的一维分离模式的分辨率和峰容量有限 .多维柱联用技术比一维分离有更高的分辨率和峰容量 [2 ] ,便于和 MS直接联用 [3,4 ] ,易于实现自动化 .目前 ,有关 2 D- CE的报道相对较少 [5～ 7] ,我们初步实现了将 2 D- PAGE由平板转移到毛细管中[6 ,7] ,但凝胶柱的制作烦琐 ,存在交叉污染 ,不能与 MS直接联用 .本文用微透析中空纤维膜为接口构建了毛细管等电聚焦 ( …     

毛细管电泳技术在单克隆抗体药物分析中的应用

单克隆抗体药物在生物制药行业占有重要地位,是生物医药领域发展的主要方向。因此,单克隆抗体药物的质量控制已成为全球生物制药企业及法规机构关注的热点,对单克隆抗体药物精确表征的需求日益增加。毛细管电泳技术具有分离效率高、分析速度快、分离模式多、样品用量少等特点,已成为单克隆抗体药物分析和质量控制的重要手段。该文对毛细管凝胶电泳、毛细管等电聚焦、毛细管区带电泳等模式在单克隆抗体药物的纯度分析、等电点测定、电荷异质性分析和N-寡糖分析的应用进行综述,以期为国内单克隆抗体研究开发和生产的企事业单位提供技术参考。     

一种电驱动高倍率、连续、无损伤浓缩装置的研究

提出使用膜聚焦浓缩方法并建立电驱动高倍率、连续、无损伤装置.分别以溴酚蓝、荧光素钠溶液为浓缩对象进行测试.该实验装置达近2个数量级浓缩效果,并可根据需要放大,成为有规模产量的生产设备,也可微型化,成为结构简单小巧、可以装配到电泳、色谱、质谱或光谱等分离分析仪器的零部件,或集成在芯片上,成为芯片实验室的预浓缩元件.     

基于移动中和界面电泳酸碱滴定的新原理及装置

基于移动中和界面(MNB)概念,研制了一种基于电泳酸碱滴定(EABT)的新装置.其核心技术是在电泳管中建立MNB,通过对电泳管中的MNB在一定时间内迁移距离的测定,推测酸或碱的浓度.EABT实验表明:HCl浓度的对数值与MNB移动距离存在线性关系.此关系可用于检测未知酸的浓度；不同酸碱指示剂对EABT方法没有明显影响,这能有效避免在容量分析法中不同指示剂对判定终点断滴所造成的误差；EABT具有良好的精密性和稳定性,日内差和日间差的RSD值分别小于1.6％和3.8％.     

A Novel Method for Studying the interaction of Macromolecule with Small Molecule by Means of Affinity Capillary Electrophoresis

IntroductionThedeterminationofbindingconstantsplaysanimportantroleforthestudiesofbiomoleculerecognition.Comparedwithtraditionalmethodstheaffinitycapillaryelectrophoresis(ACE)hasadvantagesofsmallsampleconsumption,andobservingtheinteractioninfreesoluti...     

A Computer Method for Predicting the Electrophoretic Mobility of Peptides

A computer model is presented for the prediction of the electrophoretic mobilities of peptides from physical constants derived from their amino acid sequences. The model assumes that the electrophoretic mobility can be represented by a product of four functions according to the relation:  l_comp= l(L)w(W)q(Q)c(CC), where L (a length parameter) is represented by the number of amino acid residues of the peptide, W (a width parameter) is represented by the average residue mass, Q = the charge of the peptide, and CC = the position of the center of charge relative to the center of mass. The model was used to calculate the electrophoretic mobilities of peptides in a 50 mM phosphate buffer at pH 2.5. Sixty-four test peptides ranging in size from 2 to 39 amino acid residues were used for this study. The calculated mobilities show excellent correlation with experimental measurements with a correlation coefficient greater than 0.98.     

配位作用毛细管电泳方法研究

研究了一种以过渡金属离子为添加剂,基于配位相互作用的毛细管电泳技术。选择7种芳香胺类化合物作为分析对象,考察了金属离子种类和浓度、运行介质酸度、电压对配位毛细管电泳的影响。通过对邻苯二胺变质样品的鉴别以及实际合成药磺胺类药物的分离,考察了这种毛细管电泳分离技术的分离能力,效果令人满意。应用于颠茄磺苄啶片中磺胺甲噁唑(SMZ)的定量测定时,SMZ的检出限为0.12 mmol/L;线性范围为0.25~10 mmol/L(r2=0.9917);平均加样回收率为97.8%,表明所建立的配位作用毛细管电泳(C ICE)方法,具有稳定、可靠的定性和定量分析能力。     

Particle electrophoresis and dielectrophoresis in curved microchannels

Studies of particle electrophoresis have so far been limited to primarily theoretical or numerical analyses in straight microchannels. Very little work has been done on particle electrophoretic motions in real microchannels that may have one or multiple turns for reducing the devices size or achieving other functions. This article presents an experimental and numerical study of particle electrophoresis in curved microchannels. Polystyrene microparticles are found to migrate across streamlines and flow out of a spiral microchannel in a focused stream near the outer wall. This transverse focusing effect arises from the dielectrophoretic particle motion induced by the nonuniform electric field intrinsic to curved channels. The experimental observations agree quantitatively with the numerical predictions.     

Capillary Electrophoresis: Methods and Scope

Capillary electrophoresis is a new analytical technique that has seen a great upswing in recent years, because this adaptation of electrophoresis may be automated and simple, direct quantification is possible. As in conventional electrophoresis, samples with a large range of molecular weights, from inorganic ions to biopolymers such as DNA and proteins, can be separated. Uncharged molecules can also be separated when micelle-forming detergents are added to the buffer; the distribution mechanism that then comes into play increases the separation efficiency of this micellar electrokinetic chromatography. Chiral additives such as cyclodextrins make the separation of enantiomers possible. The rapid increase in the popularity of capillary electrophoresis is reflected in instrument sales and the growing number of scientific publications dealing with the method, and it seems that the future of this technique is assured.     

Electrophoresis versus electrochromatography

In separation techniques, such as Liquid Chromatography and Capillary Zone Electrophoresis, separation is performed on the basis of differences in velocity of the various separands, making use of differences in k′ and/or effective mobility. While in chromatography the flow of the eluent is elementary, in electrophoretic techniques the electroosmotic flow is generally suppressed in order to avoid disturbing of the sample zone boundaries, which migrate with a maximal velocity of 10^?3 m s^?1. This holds especially for isotachophoretic separations, where separands migrate in consecutive zones with minimal detectable lengths of about 0.1 mm. If electroosmotic flow is applied as a transport mechanism, using capillaries as small as about 50 μm, linear velocities of the liquid flow can reach about 2 × 10^?3 m s^?1. Especially for ionic species with a low effective mobility, this velocity can be a multiple of the electrophoretic migration velocity in the separation compartment. Therefore, anionic, non-ionic, and cationic separands can migrate in the same direction. Depending on whether repulsive or attractive forces are operative, the electrophoretic separation power can be counteracted or favored. The separation mechanisms making use of (quasi)stationary phases are studied. Plotting the chromatographic behavior versus the electrophoretic shows transition areas to exist between the “purely” electrophoretic techniques and the “purely” chromatographic techniques. It must be stated that most of the recent publications in CZE, especially those with very narrow bore capillaries, can be allocated to the transition areas, sometimes with a strong chromatographic retention component.