模拟抗原(Ac-NK16-Ahx-3)免疫反应抗体ELISA检测方法研究

The Research of the Mimotop(Ac-NK16-Ahx-3)'s Immune rResponse Antibody with ELISA Test Method

为满足实验室单克隆抗体的制备研究,本实验特对实验室常用的制备单抗的模拟抗原(Ac-NK16-Ahx-3)进行ELISA间接法检测,进而建立一种高效的检测方法。通过对不同稀释度待测抗体的免疫吸附测定,检测出它的最终灵敏度为0.078mg/mL,最佳二抗稀释倍数为300倍,其最佳拟合曲线为y=-0.0732x+0.6875,R2=0.8619。并且,通过统计学方法进行统计分析,确定出各实验组的相关性及变异系数,从而排除实验中的操作误差,最终可确定最佳的实验条件,并保证实验的准确性。

This experiment is to research the mimotop （Ac-NK16-Ahx-3） which always conduet the preparation of McAb for the lab by ELISA to study and find an effectual method. In this experiment, by using ELISA to study anti- body in different dilution, the final sensitivity is 0.078ug/mL and the best dilution multiple of the secondary antibody is 300. And the best fitted curve is y= - 0.0732x＋0.6875, R2=0.8619. In addition, the statistic analysis about the result of the parallel experiment was conducted by using statistical methods and each experiment＇ s relativity and RSD.was definited.Finally the best experimental condition was determined and the accuracy of the experiment was assured.