间接抗体夹心酶联免疫吸附法测定人红细胞生成素(EPO)体外活性

目的　介绍一种经济、实用的红细胞生成素(EPO)体外活性检测方法—间接抗体夹心酶联免疫吸附法。方法　采用捕捉抗体(单克隆抗体:鼠抗EPO)包被酶标板,然后用封闭液封闭酶标板,加入稀释成合适浓度的EPO样品进行温育,再加入EPO第二抗体(多克隆抗体:兔抗EPO)温育,然后加入酶连抗体(HRP -羊抗兔)温育,最后加入底物缓冲液显色,用酶标仪读数,计算出EPO体外效价。结果　该方法重复性好(CV <5 %),与ELISA试剂盒比较,两法测定的结果相符( P >0 . 5 )。结论　该方法是一种经济、实用、准确、可靠的红细胞生成素(EPO)体外活性检测方法,与EPOELISA试剂盒比较,大大降低了实验费用

Determination of Erythropoietin (EPO) potency in vitro with indirection antibodies sandwich in enzyme linked immunosorbent method

Objective An economic and practical determination method of EP O potency in vitro with indirection antibodies sandwich in enzyme linked immunos orbent method was reported.Methods The microplate was coated with murine against EPO monoclonal antibody,then seal the microplate with seal ing solution,add specimens diluted to suitable concentration to microplate,inc ubate for some time,add the rabbit against EPO polyclonal antibody,incubate f or some time,then add HRP-goat against rabbit antibody,incubate for some time,a d d substrate buffer,incubate for some time,determine the optical density(O.D) us ing a microplate reader and count the EPO potency in vitro.Results The method has a good repetivity(CV<5%).The result is in accordance wit h the epo ELISA kit(P>0.5).Conclusion The method was e co nomical,practical for the determination of EPO potency in vitro with good repetivity and accuracy,the test cost can be reduced largely compared with the epo ELISA kit.