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压电免疫传感技术快速筛检花生中过敏原

     

摘要

Objective The study was to establish a new method for the rapid screening of peanut allergen by piezoelectric immunosensing technique. Methods Peanut major allergen proteins were isolated and puri-fied. Peanut antibodies were gotten by immunizing New Zealand white rabbits. The potency, IC50 equivalents were determined. The use of self-assembly of a piezoelectric sensor coated peanuts immune antibodies, making the standard curve to determine the sensitivity, recovery and reproducibility. The sensitivity, recovery and reproducibility were detected by the standard curve of using self-assembly piezoelectric immunosensor coated peanut antibody. Results The purity of peanut allergen protein was 90.7%, and the most suitable concentration of coated peanut antigen was 0.1μg/mL. There is a good linear relationship in range of 1μg/mL to 316μg/mL by self-assembly piezoelectric sensors to detect peanut allergen concentration. The de-tection sensitivity was 0.1μg/mL. Recovery rate was between 92.3%~113.4%and repeatability coefficient of variation was 6.04%. Conclusion Compared with other optical methods, this method has a higher sensitiv-ity and also has other advantages such as rapid detection, online detection, low costs, with simple equipment, and it can be widely applied.%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。

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