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目的:制备微囊藻毒素-LR(简称MC-LR)多克隆抗体,并将其应用于电化学免疫传感器对MC-LR的测定。方法:通过对新西兰大白兔免疫自制免疫原MC-LR-KLH,制备多抗。将所得抗体应用于电化学免疫传感技术,基于L-半胱氨酸自组装技术和纳米金吸附作用相结合的方法固定MC-LR抗体,采用循环伏安法(CV)以及交流阻抗法(EIS)研究了该抗体在修饰电极上的电化学性质。结果:间接ELISA表明纯化后的抗体效价能达到6.4×104;固定包被抗原MC-LR-BSA,采用间接竞争ELISA测定标准曲线,定量检测区间为0.05～4μg/L,半数抑制率为0.68μg/L。将所制备抗体应用于电化学研究中,成功地建立了阻抗型免疫传感器检测MC-LR。该免疫传感器显示出响应速度快,15min内完成测定;灵敏度高,达到1.82×10-2μg/L;线性范围广(0.05～200μg/L)。结论:制备的MC-LR抗体具有效价高,特异性强,应用于电化学免疫传感检测方法的研究中,体现出良好的发展前景。
Objective: To prepare microcystin-LR (MC-LR) polyclonal antibody and apply it to the determination of MC-LR by electrochemical immunosensor. Methods: New Zealand white rabbits immunized with self-made immunogen MC-LR-KLH, polyclonal antibody was prepared. The obtained antibody was applied to electrochemical immunosensor technology, MC-LR antibody was immobilized on the basis of L-cysteine self-assembly technology and adsorption of gold nanoparticles. Cyclic voltammetry (CV) and AC impedance method EIS) studied the electrochemical properties of the antibody on the modified electrode. Results: Indirect ELISA showed that the titer of purified antibody reached 6.4 × 104. The standard curve of indirect ELISA was used to immobilize antigen coated MC-LR-BSA. The quantitative detection range was 0.05-4 μg / L, and the half inhibition rate was 0.68 μg / L. The prepared antibody was used in electrochemical studies, the successful establishment of an impedance-type immunosensor MC-LR detection. The immunosensor showed a fast response and completed the assay within 15 min. The sensitivity was high at 1.82 × 10-2 μg / L and the linear range was wide (0.05-200 μg / L). CONCLUSION: The prepared MC-LR antibody has high titer and specificity and is used in the research of electrochemical immunosensing detection method, which shows a good prospect of development.