MTS染色法测定重组人肿瘤坏死因子相关凋亡诱导配体的生物学活性  

Determination of Bioactivity of Recombinant Human TNF-related Apoptosis-inducing Ligand by MTS Staining

作　　者：陈红霞 于玉根 张慧娟 黄立勤 

机构地区：[1]深圳新鹏生物工程有限公司,广东深圳518057

出　　处：《中国生物制品学杂志》2011年第1期99-101,共3页Chinese Journal of Biologicals

摘　　要：目的 建立稳定可靠的重组人肿瘤坏死因子相关凋亡诱导配体(Recombinant human TNF-related apoptosis-induc-ing ligand,rhTRAIL)生物学活性检测方法——MTS染色法。方法采用人非小细胞肺癌细胞(NCI-H460)凋亡-MTS染色法对3批rhTRAIL供试品进行检测,采用四参数回归法计算供试品效价;并对该方法的专属性、精密性和准确性进行验证。结果 3批供试品(20071101、20071102、20071103)的效价分别为7.3×104、8.1×104和9.3×104 U/ml,相关系数(R2)分别为0.998、0.996和0.998。rhTRAIL抗体能完全阻断rhTRAIL与NCI-H460细胞间的应答反应。该方法专属性较好,不同检测板间、不同加样位置、不同工作日间和不同试验人员间检测结果的平均变异系数分别为12%、26%、24%和7%,平均加样回收率为114%。结论 MTS染色法测定rhTRAIL的生物学活性具有较高的准确性和可信度,适用于rhTRAIL生物学活性的常规检测。Objective To develop a stable and reliable MTS staining method for determination of bioactivity of recombinant human TNF-related apoptosis-inducing ligand（rhTRAIL）.Methods Three batches of test samples of rhTRAIL were determined by induction of human non-small cell lung cancer NCI-H460 cell apoptosis and MTS staining,and calculated for potency by four-parame-ter regression method,based on which the specificity,precision and accuracy of the developed method were verified.Results The potencies of the three batches of test samples,with lot numbers of 20071101,20071102 and 20071103,were 7.3 × 104,8.1 × 104 and 9.3 × 104 U / ml,while the correlation coefficients（R2）were 0.998,0.996 and 0.998,respectively.The antibody against rhTRAIL completely blocked the reaction of rhTRAIL with NCI-H460 cells.The method showed high specificity,of which the mean variation coefficients of determination results using various test plates,at various sample loading sites,on various dates and by various personnel were 12%,26%,24% and 7% respectively,with a mean recovery rate of 114%.Conclusion MTS staining showed high accuracy and reliability and was suitable for the routine determination of bioactivity of rhTRAIL.

关 键 词：TNF相关凋亡诱导配体 生物学活性 MTS 

分 类 号：Q511[生物学—生物化学] R392-33[医药卫生—免疫学]