亲和毛细管电泳技术及其应用

对近几年新发展起来的亲和毛细管电泳技术（ACE）的原理、分类及方法作了简要介绍,着重介绍了亲和毛细管区带电泳、毛细管亲和凝胶电泳、胶束电动色谱中的亲和电泳、亲和毛细管等电聚焦、亲和探针毛细管电泳等过程和方法。对ACE在分子生物学、生物化学中的应用及该技术在亲和常数测定、核酸片段识别、竞争免疫分析、药物先导化合物的筛选等方面的应用也作了介绍。     

亲和膜色谱技术研究进展

对一种崭新的生物大分子分离技术－－亲和膜色谱技术做了全面的评述,介绍了亲和膜色谱分离的过程,评价了亲和膜基质材料活化、改性方法、给出了配基间隔臂的选择原则,概括了亲和膜色谱理论研究的进展,并对亲和膜色谱技术进行了展望。     

膜亲和色谱的现状、发展和应用

文章对膜亲和色谱的原理、特点、设备、过程和发展情况做了介绍，并对亲和膜在整个膜分离技术中的地位、所占的比重及市场预测做了述评，对膜亲和色谱与其它色谱分离技术的优缺点进行了比较。重点介绍了制备亲和膜的材料，活化方法，问隔臂和配基的种类、选择和共价键合方法，配基和配合物产生亲和作用的机理及解离过程和方法。并对膜亲和色谱在酶、蛋白质、核糖核酸等生物大分子纯化分离方面的应用情况做了述评。     

硼亲和材料的研究进展

硼亲和材料作为一种选择性分离富集顺式二羟基生物分子的功能性材料,近十几年里得到了蓬勃发展。在常规硼亲和材料的基础上,一些结构新颖、功能独特的材料逐渐出现并得到了广泛应用。本文介绍了硼亲和材料的研究进展,并总结了该类材料在硼亲和分离富集、疾病诊断、适配体筛选、药物传递和生物传感等方面的应用,最后对硼亲和材料的发展前景进行了展望。     

膜亲和色谱的现状、发展和应用

文章对膜亲和色谱的原理、特点、设备、过程和发展情况做了介绍，并对亲和膜在整个膜分离技术中的地位、所占的比重及市场预测做了述评，对膜亲和色谱与其它色谱分离技术的优缺点进行了比较。重点介绍了制备亲和膜的材料，活化方法，间隔臂和配基的种类、选择和共价键合方法，配基和配合物产生亲和作用的机理及解离过程和方法。并对膜亲和色谱在酶、蛋白质、核糖核酸等生物大分子纯化分离方面的应用情况做了述评。     

肽配基纳米亲和载体定向固定化抗体

抗体经常被固定在某一固相载体表面用于免疫检测分析,但通常得到的是随机固定化的抗体,因而会影响抗体的活性。本文开发了一种新型的亲和载体用于抗体的定向固定化。即将与抗体IgG的Fc片段具有亲和作用的肽配基偶联于pGMA纳米球上,制备亲和载体用于吸附抗体实现抗体的定向固定化。首先利用乳液聚合法制备了粒径为110nm的pGMA纳米球,再以EDMA为交联剂并且在纳米球表面氨基化,最后将与抗体IgG的Fc片段具有亲和作用的肽配基HWRGWV偶联于纳米球表面,制备得到肽配基纳米球亲和载体。然后,考察了亲和载体对猪IgG的吸附行为。进一步,利用等温滴定微量量热仪研究了该纳米球亲和载体与IgG之间的相互作用热力学。结果表明,肽配基亲和载体对IgG的特异性吸附作用明显高于肽配基修饰前的载体。亲和载体与IgG的亲和作用主要贡献是疏水相互作用;而IgG与氨化纳米球载体的吸附作用则主要通过静电作用和氢键作用。所制得的纳米球亲和载体可用于进一步开发高效率、高灵敏度的临床免疫检测方法。     

以寡聚组氨酸为配基的高效亲和色谱研究

亲和色谱在所有色谱分离中选择性最高.以组氨酸为配基的亲和介质(Affinitymedia)对免疫球蛋白IgG具有亲和力,显示了良好的分离效果[1];以组氨酸为配基的亲和色谱分离纯化IgG的机理[2]以及组氨酸亲和色谱分离IgG亚基也已有报道[3].组氨酸配基亲和色谱法具有简单、有效和价廉等特点,在免疫化学、临床分析及血液制品的生产等方面均有良好的应用前景.本文分别以含1,3,5个组氨酸残基的小肽为配基,合成了系列亲和介质,并对其亲和色谱行为进行了评价,研究了不同链长的寡聚组氨酸与IgG的亲和相互作用.1　实验部分1.1　固相肽合成　采用Fmoc肽合…     

亲和膜用于去除医药及人血清白蛋白溶液中的内毒素(英文)

 以纤维素膜为基质材料 ,制备了 3种用于去除内毒素的亲和膜 ,分别为壳聚糖亲和膜 (KFCC5 17) ,疏水阳离子亲和膜 (KFCG316 )和金属螯合物亲和膜 (KFCG40 2 )。研究了亲和膜对内毒素的吸附容量及在内毒素去除方面的应用 ,结果表明这 3种亲和膜都可以用于多种溶液中内毒素的去除。考察了离子强度、pH值以及流速对去除效果的影响 ,并分析了原因。KFCC5 17,KFCG316 ,KFCG40 2都可用于人血清白蛋白溶液中内毒素的去除 ,KFCC5 17还可用于医药制剂如氢化可的松、葛根素、盐酸丁卡因和右旋糖苷 40葡萄糖注射液中内毒素的去除。     

亲和色谱仿生配基的筛选和设计

利用选择性好、稳定性高、价格低廉的仿生配基作为亲和色谱的功能基团,实现对目的蛋白质的亲和分离,可以弥补常规的单克隆抗体和其他天然蛋白质配基在价格、稳定性方面的不足,从而拓展亲和色谱技术的应用范围。本文综述了亲和色谱仿生配基在筛选和设计方面的研究进展,重点介绍了组合方法和基于结构的仿生配基的设计及其应用。     

量子点标记链霉亲和素及其生物活性检测

选用无机盐为前驱体,在水相中合成CdTe量子点,并用此量子点标记链霉亲和素,通过SephadexG-100层析分离纯化量子点标记的链霉亲和素,采用磁颗粒标记的链霉亲和素与量子点标记的链霉亲和素竞争结合辣根过氧化酶标记的生物素,即酶联免疫竞争抑制分析法检测链霉亲和素标记量子点后的生物活性,计算约70.3%的链霉亲和素标记到量子点上,且具有生物活性。每毫克量子点大约可偶联0.14 mg的链霉亲和素。采用荧光光谱研究量子点标记前后的荧光变化,标记后量子点的最大发射波长蓝移了8 nm,而发射光谱的半峰宽基本不变,说明量子点与链霉亲和素结合后粒子没有团聚,分散性好。     

Determination of binding constants by affinity chromatography

This review summarizes developments in the use of affinity chromatography to characterize biospecific interactions in terms of reaction stoichiometry and equilibrium constant. In that regard, the biospecificity incorporated into the design of the experiment ensures applicability of the method regardless of the sizes of the reacting solutes. By the adoption of different experimental strategies (column chromatography, simple partition equilibrium, solid-phase immunoassay and biosensor technology protocols) quantitatiative affinity chromatography can be used to characterize interactions governed by an extremely broad range of binding affinities. In addition, the link between ligand-binding studies and quantitative affinity chromatography is illustrated by means of partition equilibrium studies of glycolytic enzyme interactions with muscle myofibrils, an exercise which emphasizes that the same theoretical expressions apply to naturally occurring examples of affinity chromatography in the cellular environment.     

Affinity monolith chromatography

The combined use of monolithic supports with selective affinity ligands as stationary phases has recently given rise to a new method known as affinity monolith chromatography (AMC). This review will discuss the basic principles behind AMC and examine the types of supports and ligands that have been employed in this method. Approaches for placing affinity ligands in monoliths will be considered, including methods based on covalent immobilization, biospecific adsorption, entrapment, and the formation of coordination complexes. Several reported applications will then be presented, such as the use of AMC for bioaffinity chromatography, immunoaffinity chromatography, immobilized metal-ion affinity chromatography, dye-ligand affinity chromatography, and biomimetic chromatography. Other applications that will be discussed are chiral separations and studies of biological interactions based on AMC.     

Pseudoaffinity Chromatography Using a Convective Interaction Media®-Disk Monolithic Column

Monolithic convective interaction media-disk (CIM-disk) chromatography is one of the fastest liquid chromatographic methods for the separation and purification of biomolecules due to its high mass transfer rate. In this way, all separated molecules are transported by convection into the pores of the matrix, resulting in a very fast separation due to the low mass transfer resistance of the CIM-disk. Due to the advantages of monolith technique, in recent years, CIM-disk affinity chromatography has been developed and investigated for purification of peptides, restriction enzymes, antibodies, etc. In this review, applications of monolithic affinity chromatography are discussed. The purification of restriction enzymes, polyclonal and monoclonal antibodies using a new monolithic CIM-disk system with immobilized histidine affinity chromatography are presented.     

磷酸化肽段分离富集方法研究进展

目前磷酸化肽段鉴定主要依赖于质谱技术,但磷酸化肽段的低丰度性以及来自非磷酸化肽段的干扰等因素,影响质谱的分析与鉴定。因此质谱分析前磷酸化肽段的富集,是深入研究磷酸化蛋白质组学的先决条件。该文介绍了磷酸化蛋白质组学中传统的以及新建立的一些磷酸化肽段分离富集方法的原理及优缺点,这些方法包括固相金属离子亲和色谱法(IMAC)、金属氧化亲和色谱法(MOAC)、强阳/阴离子交换色谱法(SCX/SAX)、亲水相互作用色谱法(HILIC)、静电排斥亲水相互作用色谱法(ERLIC)、化学衍生法、MALDI靶盘富集法以及多种富集方法相结合。     

Recent developments in quantitative affinity chromatography.

This review surveys developments during the past decade in the use of quantitative affinity chromatography as a means of evaluating equilibrium constants for solute-ligand and solute-matrix interactions. Topics include allowance for multivalency of the partitioning solute, removal of the myth that highly substituted affinity matrices are unsuitable for zonal quantitative affinity chromatography, adaptation of the technique to allow characterization of high-affinity interactions and the application of quantitative affinity chromatography theory to the characterization of biospecific adsorption phenomena in cellular systems.     

Fast Affinity Chromatography Using Small Particle Silica-Based Packing Materials

Summary

Affinity chromatography is one of the most powerful techniques for the purification of biologically active proteins available (for review see [1]). The ability of this method to purify proteins is based on highly specific, selective or characteristic interactions with immobilized ligands. Several advantages over traditional soft gel affinity supports have been observed with the use of small particle silica based materials for high performance affinity chromatography. These include greatly improved mass transfer properties which allow separations that are not always practical in the low performance mode, greatly reduced equilibration and isolation times, high available ligand densities, small elution volumes, excellent recovery of very small quantities of protein and high dynamic capacities. The criteria for developing a general, derivatizable, high performance support for high performance affinity chromatography are discussed. The step-by-step examination of these criteria and experimental evidence for determining parameters such as ligand density, non-specific adsorption and column life time for such a system are described. Chromatographic results are shown for preparative separations of (i) receptor proteins, (ii) antibodies and (iii) active enzymes.     

核酸适配体亲和色谱的研究进展

核酸适配体亲和色谱是将核酸适配体作为色谱固定相上的亲和配体的一种新型色谱技术。核酸适配体是一种可以特异性地识别目标物的寡聚核苷酸,与免疫抗体相比,核酸适配体在筛选制备、稳定性及应用等方面都显示出独特的优点。本文介绍了核酸适配体亲和色谱在小分子、蛋白质和细胞的分离和分析中的应用,对核酸适配体亲和色谱的研究现状和发展前景进行了综述。     

应用前沿亲和色谱研究分子之间相互作用及其应用

分子间的相互作用是一切生物或化学变化的基础。前沿亲和色谱作为通用性的研究分子间相互作用力的工具,利用它可得到分子之间相应的解吸常数K_d,并可对各种配体的亲和力大小进行排序。本文介绍了前沿亲和色谱的工作原理、前沿亲和色谱柱的构建,以及在药物筛选,生物相关领域的应用。与其他研究分子间相互作用的技术作了相应的比较。     

Recent stationary phase-based fractionation strategies in proteomic analysis

In this article, we review stationary phase-assisted sample prefractionation in proteomic analysis in 2019–2022 period. Applications are grouped according to the mode of retention used in the prefractionation step prior to the final analysis by low-pH reversed-phase liquid chromatography-tandem mass spectrometry. Online or offline instrumental configurations are also commented on, with a highlight on novel online platforms. From the articles reviewed in this period, the order of popularity of chromatographic modes for sample fractionation is affinity chromatography > size exclusion chromatography > hydrophilic interaction chromatography > high-pH reversed-phase liquid chromatography > ion exchange chromatography.     

Applications of silica supports in affinity chromatography

The combined use of silica-based chromatographic supports with immobilized affinity ligands can be used in many preparative and analytical applications. One example is the use of silica-based affinity columns in HPLC, giving rise to a method known as high-performance affinity chromatography (HPAC). This review discusses the role that silica has played in the development of affinity chromatography and HPAC and the applications of silica in these methods. This includes a discussion of the types of ligands that have been employed with silica and the methods by which these ligands have been immobilized. Various formats have also been presented for the use of silica in affinity chromatographic methods, including assays involving direct or indirect analyte detection, on-line or off-line affinity extraction, and chiral separations. The use of silica-based affinity columns in studies of biological systems based on zonal elution and frontal analysis methods will also be considered.