间接酶联免疫法检测麻疯树核糖体失活蛋白

Determination of curcin by indirect enzyme-linked immunosorbent assay

建立了基于单克隆抗体的间接非竞争酶联免疫法用于检测麻疯树核糖体失活蛋白的含量.优化单克隆抗体最佳工作浓度为312.5ng/mL,辣根过氧化物酶标记山羊抗小鼠IgG最佳工作浓度为200ng/mL.该方法的线性范围为25～300ng/mL,线性回归方程为y=0.0024x＋0.0322,相关系数R^2=0.9985,定量检测限25ng/mL,定性检出限3.0336ng/mL.试验证明该方法准确度较高,且具有较好的重复性和特异性,可以用于实际检测麻疯树种仁核糖体失活蛋白的含量.

An indirect non-competitive enzyme-linked immunosorbent assay （ELISA） was established to detect the content of curcin in Jatropha seeds by using monoclonal antibody. The best working concen- tration of monoclonal antibodies and peroxidase-conjugated affinipure goat anti mouse IgG were 312. 5 ng/mL and 200 ng/mL respectively. The linear range of the method was 25～300 ng/mL. The linear re- gression equation was y = 0. 0024x ＋ 0. 0322 with the linear correlation coefficient R^2 = 0. 9985. The quantification limit was 25 ng/mL meanwhile the determination limit was 3. 0336 ng/mL. Repeatability test and specificity test proved that the method has good repeatability and specificity. This method is suitable for qualitative determination of curcin.