Jones C, Patel A, Griffin S, Martin J, Young P, O'Donnell K, Silverman C, Porter T, Chaiken I
Department of Protein Biochemistry, SmithKline Beecham, King of Prussia, PA 19406, USA.
J Chromatogr A. 1995 Jul 14;707(1):3-22. doi: 10.1016/0021-9673(95)00466-z.
Molecular recognition guides the selective interaction of macromolecules with each other in essentially all biological processes. Perhaps the most impactful use of biomolecular recognition in separation science has been in affinity chromatography. The results of the last 26 years, since Cuatrecases, Wilchek and Anfinsen first reported the purification of staphylococcal nuclease, have validated the power of biomolecular specificity for purification. This power has stimulated an explosion of solid-phase ligand designs and affinity chromatographic applications. An ongoing case in point is the purification of recombinant proteins, which has been aided by engineering the proteins to contain Affinity-Tag sequences, such as hexa-histidine for metal-chelate separation and epitope sequence for separation by an immobilized monoclonal antibody. Tag technology can be adapted for plate assays and other solid-phase techniques. The advance of affinity chromatography also has stimulated immobilized ligand-based methods to characterize macromolecular recognition, including both chromatographic and optical biosensor methods. And, new methods such as phage display and other diversity library approaches continue to emerge to identify new recognition molecules of potential use as affinity ligands. Overall, it is tantalizing to envision a continued evolution of new affinity technologies which use the selectivity built into biomolecular recognition as a vehicle for purification, analysis, screening and other applications in separation sciences.
在几乎所有生物过程中，分子识别引导着大分子之间的选择性相互作用。生物分子识别在分离科学中最具影响力的应用或许是在亲和色谱方面。自夸特雷克阿斯、威尔切克和安芬森首次报道葡萄球菌核酸酶的纯化以来，过去26年的研究结果证实了生物分子特异性在纯化方面的强大作用。这种强大作用推动了固相配体设计和亲和色谱应用的迅猛发展。一个正在进行的例子是重组蛋白的纯化，通过对蛋白质进行工程改造使其包含亲和标签序列，如用于金属螯合分离的六组氨酸和用于通过固定化单克隆抗体进行分离的表位序列，这对重组蛋白的纯化起到了帮助作用。标签技术可适用于平板分析和其他固相技术。亲和色谱的发展也推动了基于固定化配体的方法来表征大分子识别，包括色谱法和光学生物传感器方法。而且，诸如噬菌体展示和其他多样性文库方法等新方法不断涌现，以鉴定有潜力用作亲和配体的新识别分子。总体而言，设想新的亲和技术持续发展是很诱人的，这些技术利用生物分子识别中固有的选择性作为分离科学中纯化、分析、筛选及其他应用的手段。
