Winzor D J, Munro P D, Cann J R
Department of Biochemistry, University of Queensland, Brisbane, Australia.
Anal Biochem. 1991 Apr;194(1):54-63. doi: 10.1016/0003-2697(91)90150-r.
In an investigation of the problem of determining kinetic parameters for the interaction of a solute with immobilized ligand sites on an affinity matrix, a combination of experimental studies and numerical simulations of frontal chromatography of methyl orange on Sephadex G-25 has yielded a simpler method than existing procedures for characterizing solute-matrix kinetics. A significant change in approach has entailed the direct evaluation of the kinetic contribution to boundary spreading from the flow-rate dependence of boundary variance under conditions of concentration-independent chromatographic migration (linear kinetics). This kinetic contribution is then interpreted in terms of an experimentally more appropriate form of a quantitative relationship for diffusion-free chromatographic migration (H. W. Hethcote and C. DeLisi, 1982, J. Chromatogr. 240, 269-281). Finally, the results of numerical simulations of concentration-dependent chromatographic migration (Langmuir kinetics) have indicated that rate constants should also be determinable under these conditions by extrapolation of their apparent values obtained by the above procedure to infinite dilution.
在一项关于确定溶质与亲和基质上固定化配体位点相互作用动力学参数问题的研究中，对甲基橙在葡聚糖凝胶G - 25上进行前沿色谱的实验研究与数值模拟相结合，产生了一种比现有程序更简单的方法来表征溶质 - 基质动力学。方法上的一个重大变化是在浓度无关色谱迁移（线性动力学）条件下，根据边界方差对流速的依赖性直接评估边界扩展的动力学贡献。然后根据无扩散色谱迁移的定量关系（H. W. Hethcote和C. DeLisi，1982，《色谱杂志》240，269 - 281）的一种在实验上更合适的形式来解释这种动力学贡献。最后，浓度依赖性色谱迁移（朗缪尔动力学）的数值模拟结果表明，在这些条件下，速率常数也应该可以通过将上述程序获得的表观值外推到无限稀释来确定。
