Chaiken I M
J Chromatogr. 1986 Apr 11;376:11-32. doi: 10.1016/s0378-4347(00)80821-x.
Measuring macromolecular and cellular interactions remains central to the study of recognition in biology and its application in biotechnology. Analytical affinity chromatography provides a versatile methodology to detect and quantitate such interactions. Both zonal and frontal elution approaches have been developed, essentially in parallel, for analytical affinity chromatography. A close quantitative relatedness of chromatographically obtained equilibrium constants and analogous constants determined fully in solution has been found for a growing number of proteins. This consistently observed correlation has formed the basis for extending theoretical treatments in order to evaluate not only monovalent molecular systems of varying types but also multivalently interacting macromolecules, including those which exhibit cooperativity. The potential to measure chemical rate constants by affinity chromatography also has been recognized, and experimental tests of the available theory are being made. As a micromethod, the quantitative use of affinity chromatography has important applicability for biochemical analysis of an increasing array of biologically active molecules being discovered and isolated but available in only relatively small amounts. Analytical affinity chromatography thus provides a means to use matrix--mobile interactant systems to study mechanisms of biomolecular interactions and therein to attain an understanding of such interactions which often is not easily achieved by solution methods alone.
测量大分子和细胞间的相互作用仍然是生物学中识别研究及其在生物技术中应用的核心内容。分析亲和色谱法提供了一种通用的方法来检测和定量此类相互作用。区域洗脱和前沿洗脱方法基本上是并行发展起来用于分析亲和色谱的。对于越来越多的蛋白质，已发现色谱法获得的平衡常数与在溶液中完全测定的类似常数之间存在密切的定量相关性。这种一直观察到的相关性为扩展理论处理奠定了基础，以便不仅评估不同类型的单价分子系统，还能评估多价相互作用的大分子，包括那些表现出协同性的大分子。通过亲和色谱法测量化学速率常数的潜力也已得到认可，并且正在对现有理论进行实验测试。作为一种微量方法，亲和色谱法的定量应用对于越来越多被发现和分离但数量相对较少的生物活性分子的生化分析具有重要的适用性。因此，分析亲和色谱法提供了一种利用基质 - 流动相互作用剂系统来研究生物分子相互作用机制的手段，并借此了解此类相互作用，而这往往仅靠溶液方法是不容易实现的。
