吖啶酯化学发光测量参数和睾酮抗体工作浓度的最佳化

OPTIMAL PARAMETERS OF MEASURING TESTOSTERONE-3-ACRIDINIUM ESTER AND ANTIBODY TITER FOR CHEMILUMINESCENCE IMMUNOASSAY

１０－甲基吖啶－９－羧酸［４－（Ｎ－睾酮－３－羧甲氧基肟）－氯乙基］苯酯（Ｔｏ－３－ＡＥＰＭＡＣ）发光测量的时间参数在低浓度区域测量十分重要．本文报道在０．００～０．２１ｓ间有一占发光总面积４．１０±２．２１％（±ｓ）的溶剂峰；峰发光的起始和终点在０．３２～３．９１５间，峰时间参数位于０．４３～２．２２ｓ间，反应速率分析也证实时间参数的实验结果．选择０．３０ｓ延迟时间和１０ｓ积分时间的化学发光积分参数测量既可排除溶剂发光的干扰，又可较精密地采集到Ｔ０－３ＡＥＰＭＡＣ的全部发光信号。而单位时间的发光峰高测量因随剂量增加峰时间参数延长不能满足建立超微量化学发光免疫化学分析测量的要求．本文根据化学发光免疫化学高灵敏和高精密的分析要求，通过抗体稀释曲线极值的优选、最大反应变化的标定和最小反应标准误点分析使睾酮抗体的工作浓度最佳化，即稀释度为１：１８０００．

In order to developing sensitive and precise testoterone chemiluminescence immunoassdy,optimal paremeters of measuring acridinium ester and antibody titer must be selected.In this study,we find that time paremeter for luminescence analysis testosterone-3-carboxymethyloxime-4-(To-mehty)-acridinium-9-carboxylate(To-3AEPMAC)is very important in the low concentration.During the measurement,there is a peak of solvent in 0.00～0.21s,peak of To-3-AEPMAC in 0.43～2.22s.The luminescence reaction can be completed in 0.32～3.91s.But it is not very correction because the more To-3-AEPMAC is,the longer luminescence peak time is.These results are decided by the reaction mixing condition and reactive dynamics.The solvent peak occupies 4.10±2.21%(x±s)in all luminescence.The same result is obtained with measuing the reaction rate. It is best to choose 0.30 s delay time and 10 s integrated time for analysis the chemiluminescence characteristic.This measuring manner riot only refused the luminescence disturbance of solvent, but also obtained all signal of To-3-AEPMAC.Then it is bad to analysis the hight peak of luminescence because of varying time in peak appearing.Optimal antibody titer is measured with three methods,there are:maximum value of antibody diluted curve, maximum verying reaction and minimum reaction standard error.The first and second methods are respected for sensition,the later method is respected for precision of standard curve. The optimal antibody titer of testosterone chemiluminescence immunoassay is 1:18000.