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目的采用压电免疫传感技术快速筛检花生过敏原。方法对花生主要过敏蛋白进行分离纯化,免疫新西兰大白兔获得花生抗体,对其的效价,半数抑制浓度等值进行测定。运用自组装的压电免疫传感器包被花生抗体,制作标准曲线,确定灵敏度,回收率及重现性。结果分离纯化花生过敏原蛋白纯度为90.7%,最适的花生包被抗原浓度为0.1μg/mL,自组装的压电传感器检测花生过敏原浓度在1~316μg/mL范围内具有较好的线性关系,灵敏度为0.1μg/mL,回收率介于92.3%～113.4%之间,重现性变异系数为6.04%。结论本项目建立的压电免疫传感技术检测花生过敏原与目前较多的光学分析相比灵敏度更高,实现了快速、在线检测,成本低,仪器简单,具有广阔的应用空间。
Aim To rapidly screen peanut allergens using piezoelectric immunosensor technology. Methods To isolate and purify the major allergy protein of peanut, New Zealand white rabbits were immunized to obtain peanut antibodies, and their potency and half-value concentration were determined. A self-assembled piezoelectric immunosensor was used to coat peanut antibodies to create a standard curve for sensitivity, recovery, and reproducibility. Results The purity of the isolated and purified peanut allergen protein was 90.7%, the optimal concentration of peanut antigen was 0.1μg / mL, and the self-assembled piezoelectric sensor had good linearity in the range of 1 ~ 316μg / mL The sensitivity was 0.1 μg / mL, the recoveries ranged from 92.3% to 113.4%, and the coefficient of variation of reproducibility was 6.04%. Conclusion The piezoelectric immunosensor technology established by this project is more sensitive than traditional optical analysis methods to detect peanut allergens. It achieves rapid and on-line detection, low cost, simple instrument and wide application space.