Muller D, Yu X J, Fischer A M, Bros A, Jozefonvicz J
J Chromatogr. 1986 May 30;359:351-7. doi: 10.1016/0021-9673(86)80089-9.
Cross-linked polystyrenes modified with L-arginyl methyl ester mimic the binding site of antithrombin III and thrombin substrates. They can be used as stationary phases in high-performance affinity chromatography of thrombin. Under isocratic conditions, thrombin is strongly adsorbed on the resins when the sodium chloride concentration is lower than 0.5 M. The bound enzyme can be selectively desorbed when the salt concentration is raised to about 1.2 M. With a linear salt gradient, the specific elution of thrombin can be effected with a high recovery of its enzymatic activity. The decomposition products of thrombin, when treated with sodium dodecyl sulphate, are not retained by the stationary phase. The effects of the flow-rate and salt gradient slope on the adsorption and desorption of alpha-thrombin demonstrate the importance of kinetic parameters.
用L-精氨酰甲酯修饰的交联聚苯乙烯模拟抗凝血酶III和凝血酶底物的结合位点。它们可用作凝血酶高效亲和色谱的固定相。在等度条件下，当氯化钠浓度低于0.5 M时，凝血酶强烈吸附在树脂上。当盐浓度提高到约1.2 M时，结合的酶可被选择性解吸。采用线性盐梯度，可实现凝血酶的特异性洗脱，并使其酶活性得到高回收率。凝血酶的分解产物经十二烷基硫酸钠处理后，不被固定相保留。流速和盐梯度斜率对α-凝血酶吸附和解吸的影响证明了动力学参数的重要性。
