α_1-抗胰蛋白酶活性测定方法及其影响因素的研究  

Study of assay method of the biological activity of α1-antitrypsin and factors influencing the measurement

作　　者：陈祥松[1] 卜凤荣[1] 宿艳笋[1] 曹德英[1] 

机构地区：[1]河北医科大学药学院,河北石家庄050011

出　　处：《中国输血杂志》2006年第5期364-367,共4页Chinese Journal of Blood Transfusion

摘　　要：目的研究用发色底物法测定α1-AT生物活性时，以正常人混合血浆为参考标准的血浆稀释度范围和测定的影响因素。方法采用酶标仪测定抗胰蛋白酶与过量的胰蛋白酶反应后，剩余的胰蛋白酶与BAPNA发色反应的OD值（405nm），观察时间和温度对反应的影响，优化正常人混合血浆（30人份）稀释度范围，建立并验证血浆标准曲线，同时观察几种物质对测定的影响。结果通过优化实验．血浆的稀释倍数在1／50～1／100之间有良好的线性；PEG4000、蔗糖、Tween80／TNBP（S／D）、辛酸钠等对活性测定无明显影响，而枸橼酸钠浓度〉0．125mol／L，α1-AT生物活性测定结果偏高约20％。结论以正常人混合血浆为参考标准品，用酶标仪测定α1-AT的生物活性，快速、准确，方法稳定可靠，α1-AT制备过程中常用的几种物质，除枸橼酸钠外，对其活性测定均无影响。Objective To study the determination of biological activity of α1-AT with ehromogenic substrate and its influential factors using fresh pooled normal human plasma as relerence standard. Methods Measuring absorption value of reactions between residual trypsin and BAPNA at 405nm, after α1-AT inhibition by excess trypsin. Fresh pooled normal human plasma was used as reference standard to calculate the biological activity ofα1-antitrypsin. Resuits Good linear correlation was obtained when the plasma was diluted to 1/50 to 1/100. PEG4000, sucrose, S/D （Tween80/TNBP） and sodium caprylate did not influence the biological activity of α1-AT, butyl-AT activity was increased by about 20N when the concentration of sodium citrate was above 0. 125mol/L. Conclusion The experiment proved thatα1-AT biological activity was determined using fresh pooled human plasma as reference standard, the method is stable and reliable. Except sodium citrate, all of the materials used in the assay did not influence the deter- mination orα1-AT activity.

关 键 词：α1-抗胰蛋白酶BAPNA(N2-苯甲酰-DL-精氨酸对硝基苯胺盐酸盐) 胰蛋白酶 混合血浆 正常人 

分 类 号：R392-33[医药卫生—免疫学] Q556.9[医药卫生—基础医学]