人抗凝血酶Ⅲ生产工艺中活性检测方法的建立及初步应用

Establishment and preliminary application of active detection methods in human antithrombinⅢproduction process

目的建立和验证全自动凝血分析仪检测人抗凝血酶Ⅲ(human antithrombinⅢ,AT-Ⅲ)活性的方法。方法参照全自动凝血分析仪操作说明书进行AT-Ⅲ活性的检测,并对方法的线性、检测范围、精密度和准确度进行方法学验证。结果全自动凝血分析仪检测AT-Ⅲ活性的方法稳定,仪器自动运行标准曲线的决定系数均>0.980,定标血浆标准曲线线性范围为0.12~1.23 IU/ml,定标血浆高、中、低值和定量下限稀释样品的效价回收率均在80%~120%范围内,且高、中、低值效价回收率的相对标准偏差(relative standard deviation,RSD)均<15%,定量下限稀释样品的效价回收率的RSD<20%,供试品的批内精密度RSD均<5%,批间精密度RSD均<10%,对3批AT-Ⅲ制品及其中间品进行活性检测,每批样品分别测定3次的平均RSD均<5%。结论全自动凝血分析仪检测AT-Ⅲ活性的方法具有较好的线性、检测范围、准确度和精密度,可用于AT-Ⅲ生产工艺中活性的检测。

Objective To establish and verify the method for detecting human antithrombinⅢ(AT-Ⅲ)activity by automatic coagulation analyzer.Methods According to the operation manual ofautomatic coagulation analyzer,the AT-Ⅲactivity was detected,and the linearity and detection range,precision and accuracy of the method were verified.Results Automatic coagulation analyzer detectdAT-Ⅲactivity stably and automatically ran the standard curves with coefficients of determination all>0.980,the standard curve of the calibrator was 0.12-1.23 IU/ml,the titer recovery rate range of thehigh,middle,low value calibrator and quantitative lower limit dilution samples were 80%-120%.therelative standard deviations(RSDs)of the high,middle and low value titer recovery rates were<15%,and the RSD of quantitative lower limit dilution sample titer recovery rate was<20%,inter-batch RSDs for the test products were<5%,and the intra-batch RSDs were<10%.The average RSDs of 3 batches of ATⅢproducts and intermediates tested 3 times were all<5%.Conclusion The method of automatic coagulation analyzer for detecting AT-Ⅲactivity has good linearity,test range,accuracy and precision,which can be used for the detection of AT-Ⅲactivity in the production process.