摘要
目的研制用于丙氨酸转氨酶 (ALT)测定的参考品。方法猪心作原料 ,经硫酸铵沉淀、DEAE SephadexA 5 0、CM SephadexC 5 0、SephadexG 15 0及抗天冬氨酸转氨酶 (AST)免疫亲和色谱提纯 ,用一定的基础液稀释后分装冻干。结果提纯品ALT比活为 173U/mg蛋白 ,得率 2 2 .8% ,温度加速变性试验测得预期年失活率 ,在 - 2 0℃、4℃分别为 0 .0 5 %、0 .12 %。结论此法提纯ALT得率高 ,仅含少量AST ,冻干品稳定 ,在不同方法间与人血清有互换性 。
PurposeTo prepare a stable and intermethod properties.MethodsThe enzyme was purified from hog heart by ammonium sulfate precipitation and chromatography successively on DEAE Sephadex A50?CM Sephadex C50?Sephadex G150 and immuno affinity column of anti AST antibodies. The purified enzyme was diluted with a well defined matrix, dispensing in vials,and lyophylizing.ResultsThe purified enzyme had a specific activity of 166 kU/g protein.The yearly loss of activity for the preparation of lyophyized on the basis of accelerated degradation tests was 0.05% at -20℃, 0.12% at 4℃, respectively. Preliminary assigned value for ALT catalytic activity concentration in the reconstituted lyophilized material was 201 U/L with a 0.95 confidence interval of ± 4.9 U/L when measured by the IFCC method (without PLP) at 37℃.Conclusion The material possesses the long term stability and the comutability. It is suitable as calibrator or candidate reference material (uncertified) in clinical enzyme assay.
出处
《中国生化药物杂志》
CAS
CSCD
2002年第5期223-225,共3页
Chinese Journal of Biochemical Pharmaceutics
基金
军队医药卫生基金资助课题 (96M0 2 0 )
