猪肾γ-谷氨酸基转移酶(GGT)的提纯及其稳定性

目的 提纯活性稳定的GGT，制备用于GGT（测定的参考品。方法 用差速离心法从猪肾皮质中分离刷状缘微绒毛膜，用 Triton X－100溶解膜结合的酶，用抗GGT亲和层析柱提纯 GGT，检测提纯品的有关特性。结果 提纯品GGT比活600IU/mg蛋白，得率45.5%，未测出临床常测的酶，在不同方法间的活性比与人血清GGT相似，在特定介质中制成冻干品，经加速变性试验，在-20℃和4℃预期年失活率分别为0．02％和0．25％。结论 本法提纯GGTT速度快，得率高，几乎不含杂酶，稳定性好，在不同方法间与人血清酶有互换性，是用于制备酶参考品提纯GGT的理想方法。

Purificationn of γ - glutamyltransferase (GGT) from Hog Kidney by Immuno- affinity Chromatography and Study on Stability of purified product

[Abstract] Objective To purify stable γ glutamyltransferase (GGT) used as a reference. Methods After microvillus was isolated from hog kidney cortices by differential centrifugation, the membrane - binding enzymes were dissolved with Triton X - 100, and GGT was purified by immuno - affinity column chroma-tography with anti - GGT antibody. The purified GGT was characterized. Results The specific activity and recovery rate of purified GGT were 600IU/mg protein and 45.5% respectively. No enzyme contamination was detected out. The ratio of activities detected by various methods was close to that of GGT in human serum. The GGT was freeze - dried in a special medium, and accelerated degeneration test proved that the predicted annual relative activity loss of it were 0.02% and 0.25% at - 20癈 and 4癈 respectively. Conclusion The method is rapid, and the recovery rate of GGT is high. The GGT is pure and stable and can be used as a enzyme reference.