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目的制备罗汉果甜苷V(MogV)特异性抗血清,建立MogV含量竞争性抑制酶联免疫检测法。方法用罗汉果甜苷V-丁二酸酯-牛血清蛋白(MogV-HS-BSA)结合比约为44∶1为免疫抗原制备抗血清,用罗汉果甜苷V-丁二酸酯-人血清蛋白(MogV-HS-HSA)结合比约为64∶1为包被抗原,建立MogV含量竞争性抑制酶联免疫检测法。结果包被抗原工作浓度为1μg.mL 1,血清最佳稀释倍数为1∶8 000,在此工作浓度下,MogV浓度在1×10-4~1μg.mL 1内回归方程Y=0.158X+0.298,r=0.991 0。结论成功制备了MogV抗血清,MogV浓度在1×10-4~1μg.mL 1内对抗血清的OD(405 nm)值抑制作用呈良好的线性关系。
Objective To prepare MogV specific antiserum and establish competitive inhibition MogV ELISA. Methods The antiserum was prepared with the MogV-HS-BSA conjugate ratio of about 44:1 as the immunogen. The antiserum was extracted with Mogoside V-succinate-human serum albumin (MogV-HS-HSA) binding ratio of about 64: 1 as the coating antigen, a competitive inhibition of MogV enzyme-linked immunosorbent assay was established. Results The working concentration of coated antigen was 1μg.mL 1, the optimal dilution of serum was 1: 8000. Under the working concentration, the regression equation of MogV was 1 × 10-4 ~ 1μg.mL 1 and the regression equation was Y = 0.158X + 0.298, r = 0.991 0. Conclusion MogV antiserum was successfully prepared. The MogV concentration showed a good linear relationship with the OD (405 nm) inhibition of serum at 1 × 10-4 ~ 1μg · mL 1.