Balny C, Le Doucen C, Douzou P, Bieth J G
J Chromatogr. 1979 Jan 11;168(1):133-8. doi: 10.1016/s0021-9673(00)80701-3.
A new variety of affinity chromatography of enzymes is described which consists of building up an affinity adsorbent composed of a real substrate. The chromatography is performed at a sub-zero temperature where the turnover of the enzyme is very low or stopped. As a model system Sepharose-bound L-trialanine p-nitroanilide was for used the affinity binding of porcine pancreatic elastase, which was adsorbed to the column in a hypersaline medium at--14 degrees and eluted from the column at the same temperature using 50% (v/v) ethylene glycol. The affinity adsorbent proved to be vary specific as it did not retain trypsin, chymotrypsin and ovalbumin and retained only 20% of cytochrome c.
描述了一种新型的酶亲和色谱法，该方法包括构建一种由实际底物组成的亲和吸附剂。色谱在零下温度下进行，此时酶的周转非常低或停止。作为模型系统，使用了琼脂糖结合的L-丙氨酸对硝基苯胺用于猪胰弹性蛋白酶的亲和结合，该酶在-14℃的高盐介质中吸附到柱上，并在相同温度下用50%（v/v）乙二醇从柱上洗脱。该亲和吸附剂被证明具有很高的特异性，因为它不保留胰蛋白酶、胰凝乳蛋白酶和卵清蛋白，只保留了20%的细胞色素c。
