Winzor Donald J
Department of Biochemistry, School of Molecular and Microbial Sciences, University of Queensland, Brisbane, Qld 4072, Australia.
J Chromatogr A. 2004 May 28;1037(1-2):351-67. doi: 10.1016/j.chroma.2003.11.092.
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.
本综述总结了亲和色谱法在根据反应化学计量和平衡常数表征生物特异性相互作用方面的进展。在这方面，实验设计中纳入的生物特异性确保了该方法的适用性，而与反应溶质的大小无关。通过采用不同的实验策略（柱色谱法、简单分配平衡法、固相免疫分析法和生物传感器技术方案），定量亲和色谱法可用于表征由极其广泛的结合亲和力所控制的相互作用。此外，通过糖酵解酶与肌肉肌原纤维相互作用的分配平衡研究，阐明了配体结合研究与定量亲和色谱法之间的联系，该研究强调相同的理论表达式适用于细胞环境中天然存在的亲和色谱实例。
