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为建立酶动力学方法检测血清白细胞弹性蛋白酶(Human leukocyte elas-tase,HLE),选择高敏感底物琥珀酰-(L-丙氨酸)2-缬氨酸-对硝基苯胺。该底物在HLE催化下释放出对硝基苯胺。通过在405nm处测定产物吸光度的变化,推算出HLE的活性。结果显示,该实验的米氏常数为0.086mmol/L,最适底物浓度为2.0mmol/L,最适PH为8.0,线性范围O～300U/L;精密度试验高、中、低值分别为批内CV(％)1.25、2.60、4.80,批间CV(％)1.77、3.50、5.15;正常参考值为8～20U/L;轻、重两型胰腺炎患者血清HLE与正常对照组均有显著性差异。提示该法简捷、快速、准确、灵敏,可用于手工操作及自动化分析。
To establish an enzyme-kinetic assay for detecting human leukocyte elas-tase (HLE), a highly sensitive substrate succinyl- (L-alanine) 2-valine-p-nitroaniline was selected. The substrate releases p-nitroaniline under HLE catalysis. The activity of HLE was calculated by measuring the change in absorbance at 405 nm. The results showed that the Michaelis constant of this experiment was 0.086mmol / L, the optimum substrate concentration was 2.0mmol / L, the optimum pH was 8.0 and the linear range was 0 ~ 300U / L. The precision test of high, medium and low values (%) 1.25, 2.60, 4.80, between-run CV (%) 1.77,3.50,5.15; normal reference value is 8 ~ 20U / L; HLE in both mild and severe pancreatitis patients and normal control group There are significant differences. Prompt method is simple, fast, accurate and sensitive, can be used for manual operation and automated analysis.