Cann J R, Winzor D J
Arch Biochem Biophys. 1987 Jul;256(1):78-89. doi: 10.1016/0003-9861(87)90427-9.
A theoretical and experimental study has been made of the advancing elution profile in frontal gel chromatography of interacting systems for which the elution volume of the complex is smaller than that of the larger reactant. First, Gilbert-Jenkins theory is used to delineate the form of the elution profile from the magnitudes of the elution volumes and concentrations of reacting species. This procedure resulted in the detection of a misinterpretation of certain patterns obtained in a gel chromatographic study of the interaction between myoglobin and ovalbumin. Second, a numerical computational procedure, which incorporates both axial dispersion and concentration-dependence of species elution volumes, is used to establish the influence of these two factors on boundary shapes for such systems. Third, frontal gel chromatography on Sephadex G-75 is used to compare experimental behavior with theoretical profiles predicted for the electrostatic interaction between cytochrome c and soybean trypsin inhibitor (pH 6.8, I 0.01). Results of these experiments serve as a guide for future conduct of experiments aimed at characterization of biologically important, reversible complex formation between proteins and/or other macromolecules.
对相互作用体系在前沿凝胶色谱中的推进洗脱曲线进行了理论和实验研究，该体系中复合物的洗脱体积小于较大反应物的洗脱体积。首先，利用吉尔伯特 - 詹金斯理论，根据洗脱体积的大小和反应物种的浓度来描绘洗脱曲线的形式。这一过程导致发现了在肌红蛋白和卵清蛋白相互作用的凝胶色谱研究中获得的某些模式的错误解释。其次，采用一种数值计算程序，该程序结合了轴向扩散和物种洗脱体积的浓度依赖性，以确定这两个因素对这类体系边界形状的影响。第三，在葡聚糖凝胶G - 75上进行前沿凝胶色谱，以将实验行为与针对细胞色素c和大豆胰蛋白酶抑制剂之间的静电相互作用（pH 6.8，离子强度0.01）预测的理论曲线进行比较。这些实验结果为未来旨在表征蛋白质和/或其他大分子之间生物学上重要的可逆复合物形成的实验开展提供了指导。
