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。     

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。     

亲和膜色谱

亲和膜色谱又称亲和膜分离，其在蛋白质的分离纯化中作为一种综合性的技术出现在80年代末。亲和膜色谱主要优点是克服了颗粒状多孔载体扩散传质阻力大的缺点，代之以对流传质，这样就可以在较低的操作压和较高的流速下对目标蛋白进行快速的分离和纯化，从而缩短操作时间、提高纯化效率。本文将就近年来亲和膜色谱及其在蛋白质分离和纯化中的应用作一综述性介绍。     

染料膜亲和色谱法中膜堆的制备及应用

将染料亲和配基偶联于大孔纤维素膜上,所得亲和膜用胶粘法制成亲和膜堆,膜堆的通透性远优于通常的亲和色谱柱。装有蓝色和红色亲和膜的膜堆可分别用于人血清白蛋白和碱性磷酸酯酶的分离纯化,其中碱性磷酸酯酶可在一步操作后纯化４０倍。     

金属螯合亲和膜吸附分离与纯化溶菌酶的研究（Ⅱ）——不同金 …

采用低温氧或氨等离子体法改性聚丙烯微孔膜，基于等离子体改性膜的活化，偶联及螯合过程的机理，制备了Ｆｅ＾３＋、Ｎｉ＾２＋、Ｃｕ＾２＋等金属离子螯合亲和膜，并用于溶菌酶的吸－脱附实验。两种膜的重复吸－脱附性能相近，而Ｆｅ＾３＋螯合亲和膜基本上不能用于重复吸－脱附实验，采用补充金属螯合离子，能部分恢复亲和膜对溶菌酶的吸附量，是实现亲和膜重复使用的有效方法。     

两种固定化金属螯合复合亲和膜色谱介质制备

以纤维素滤纸为基质,通过碱处理、环氧活化、偶联亚氨基二乙酸二钠、固定化Ｃｕ＾２＋后制得了大孔纤维素亲和膜。另外,在活化后的膜上通过共价交联覆盖上琼脂糖,制得了具有类似“三明治”结构且性能优于的复合亲和膜,装柱后分别制得固定化金属螯合亲和膜色谱柱。对两种亲和膜进行牛血清白蛋白等温吸附测定显示,两者的最大吸附量分别为１．１７ｍｇ／ｃｍ＾２和１．３０ｍｇ／ｃｍ＾２,与传统的琼脂糖凝胶类介质吸附量相当,表     

高效膜色谱的发展和应用

对新兴的高效膜色谱的基质材料、膜形态、分离机理以及应用进行了综述,并对其用于对映体分离作了展望。共７６篇。     

金属螯合亲和膜吸附分离与纯化溶菌酶的研究（Ⅰ）——低温氧 …

用低温氧等离子体方法对聚丙烯微孔膜表面进行改性。扫描电镜,红外光谱和光电子能谱综合分析结果表明,膜表面上产生了－ＯＨ,－ＣＯＯＨ和Ｃ＝Ｏ等极性基团,这些基团可被活化和偶联制亲和膜。     

亲和色谱纯化超螺旋质粒DNA的研究进展

非病毒载体质粒DNA已被广泛应用于基因治疗和DNA疫苗,目前迫切需要开发其大规模制备和分离纯化方法。亲和色谱是一种高分辨率、高选择性的分离技术,在蛋白质、抗体、核酸等生物大分子的分离纯化方面显示了良好的应用前景。本文综述了亲和色谱技术在超螺旋质粒DNA分离纯化中的研究进展,总结了各种亲和色谱方法分离超螺旋质粒DNA的机理和优缺点,并展望了亲和纯化技术在质粒DNA生产和制备中的应用前景。     

对映体选择性配体交换膜的制备和DL-氨基酸的拆分

制备了一种新型光学分离膜,即带有Ｌ－脯氨酸手性选择子的交联聚乙烯醇膜,考察了ＤＬ－酪氨酸,ＤＬ－苯丙氨酸和ＤＬ－色氨酸通过膜的对映体选择性渗透性能。发现Ｌ－氨基酸优先透过膜,消旋氨基酸透过膜的对映体拆分机理类似于手性配体交换色谱方法拆分消旋氨基酸的机理,在ＤＬ－氨基酸的对映体选择性膜透过中,对映体选择性吸附着重要的作用。     

Regulating the Layered Stacking of a Covalent Triazine Framework Membrane for Aromatic/Aliphatic Separation

Membrane separation of aromatics and aliphatics is a crucial requirement in chemical and petroleum industries. However, this task presents a significant challenge due to the lack of membrane materials that can endure harsh solvents, exhibit molecular specificity, and facilitate easy processing. Herein, we present a novel approach to fabricate a covalent triazine framework (CTF) membrane by employing a mix-monomer strategy. By incorporating a spatial monomer alongside a planar monomer, we were able to subtly modulate both the pore aperture and membrane affinity, enabling preferential permeation of aromatics over aliphatics with molecular weight below 200 Dalton (Da). Consequently, we achieved successful all-liquid phase separation of aromatic/aliphatic mixtures. Our investigation revealed that the synergistic effects of size sieving and the affinity between the permeating molecules and the membrane played a pivotal role in separating these closely resembling species. Furthermore, the membrane exhibited remarkable robustness under practical operating conditions, including prolonged operation time, various feed compositions, different applied pressure, and multiple feed components. This versatile strategy offers a feasible approach to fabricate membranes with molecule selectivity toward aromatic/aliphatic mixtures, taking a significant step forward in addressing the grand challenge of separating small organic molecules through membrane technology.     

Immobilized metal affinity composite membrane based on cellulose for separation of biopolymers

Ｕｐｔｏｄａｔｅ,ｒａｐｉｄｐｕｒｉｆｉｃａｔｉｏｎｏｆｍｉｘｅｄｐｒｏｔｅｉｎｉｎｌａｒｇｅｓｃａｌｅｈａｓｂｅｅｎａｎｉｍｐｏｒｔａｎｔｒｅｓｅａｒｃｈｐｒｏｊｅｃｔｉｎｂｉｏｅｎｇｉｎｅｅｒｉｎｇｐｒｏｄｕｃｔｐｒｏｃｅｓｓｉｎｇ.Ｉｍｍｏｂｉｌｉｚｅｄｍｅｔａｌｉｏｎａｆｆｉｎｉｔｙｃｈｒｏｍａｔｏｇｒａｐｈｙ(ＩＭＡＣ)ｉｓａｎｅｆｆｉｃｉｅｎｔｍｅｔｈｏｄｅｘｔｅｎｓｉｖｅｌｙｕｓｅｄｆｏｒａｆｆｉｎｉｔｙｐｕｒｉｆｉｃａｔｉｏｎｏｆｂｉｏｌｏｇｉｃａｌｌｙａｃｔｉｖｅｓｕｂｓｔａｎｃｅ…     

Analysis of allergens in tubeimu saponin extracts by using rat basophilic leukemia 2H3 cell‐based affinity chromatography coupled to liquid chromatography and mass spectrometry

An affinity two‐dimensional chromatography method was developed for the recognition, separation, and identification of allergic components from tubeimu saponin extracts, a preparation often injected to treat various conditions as indicated by traditional Chinese medicine. Rat basophilic leukemia‐2H3 cell membranes were used as the stationary phase of a membrane affinity chromatography column to capture components with affinity for mast cells that could be involved in a degranulation reaction. The retained components were enriched and analyzed by membrane affinity chromatography with liquid chromatography and mass spectrometry via a port switch valve. Suitability and reliability of the method was investigated using appropriate standards, and then, the method was applied to identify components retained from tubeimu saponin extracts. Tubeimoside A was identified in this way as a potential allergen, and degranulation assays confirmed that tubeimoside A induces RBL‐2H3 cell degranulation in a dose‐dependent manner. An increase in Ca²⁺ influx indicated that degranulation induced by tubeimoside A is likely Ca²⁺ dependent. Coupled with the degranulation assay, RBL‐2H3 cell‐based affinity chromatography coupled with liquid chromatography and mass spectrometry is an effective method for screening and identifying allergic components from tubeimu saponin extracts.     

Application of heparinized cellulose affinity membranes in recombinant adeno-associated virus serotype 2 binding and delivery

The microporous affinity membrane based on cellulose matrices offers minimal mass-transfer effects in membrane chromatography with low nonspecific adsorption. In this research, we tested a novel application of the microporous, heparinized cellulose membranes for their affinity toward recombinant adeno-associated virus serotype 2 (rAAV2, which uses heparan sulfate proteoglycans as the primary cellular receptor) to develop a controlled, substrate-mediated viral vector delivery. We conjugated rAAV2 to an epoxy-crosslinked heparin cellulose membrane, which led to vector transduction upon cellular adhesion. When adhered, human fibroblasts exhibited proliferation kinetics similar to those on the standard polystyrene tissue-culture surface. Using fluorescent proteins as the reporter, we showed that the heparin-bound rAAV2 particles remained active and that the rAAV2-heparin binding was reversible and capable of mediating transgene delivery in cell culture. In addition, we applied the affinity membrane to adsorb unpurified rAAV2 from the crude lysate of packaging cells via the ligand–receptor binding, avoiding the use of conventional ultracentrifugation or chromatography in preparation of infectious rAAV2 for transduction. Our work explores a new application of affinity cellulose membranes in substrate-mediated viral vector delivery, which can be a useful tool in developing protocols for localized gene transfer.     

气体分离分子筛膜

本文简述了气体分离分子筛膜的研究进展,总结了气体分离分子筛膜的制备方法、分子筛膜类型、气体分离性能及催化应用,分析了气体渗透测定方法和分离机理,对分子筛膜的发展提出一些建议。     

Determination of Pharmaceuticals by Dynamic Cell Membrane Chromatography Coupled with High-Performance Liquid Chromatography

As a biological affinity chromatographic method, cell membrane chromatography (CMC) using a silica stationary phase covered with specific cell membrane has been used in screening active components. The innovation of this work is that the bioactive cell membrane and the chromatographic packing are mixed and absorbed for the first time to form the pre-column. The pre-column was placed in front of a C₁₈ column to create dynamic CMC online high-performance liquid chromatography (HPLC) system. The retention behavior and dynamic changes of pharmaceuticals were studied for this system. The results indicate that the retention time of the drug was increased and the symmetry factor reached the analytical level after the addition of the dynamic cell membrane pre-column. Therefore, the dynamic CMC coupled with HPLC system may be a potentially rapid and efficient drug analysis approach for the interaction of drug molecule and receptor on red blood cell membranes.