[关键词]
[摘要]
该研究以壳聚糖、石墨烯和纳米金为复合修饰材料固定化β-乳球蛋白抗体制备了纳米免疫传感器,建立了一种快速测定乳品中β-乳球蛋白(β-LG)过敏原的方法。用循环伏安法对该纳米免疫传感器进行了表征,以1.0 mmol/L的K3[Fe(CN)6]为探针,对试验条件进行了优化,当复合修饰液用量为5 μL,电解质pH值为7.0,抗体固载量为40 ng,孵育温度为37 ℃,孵育时间30 min时为最佳反应条件。该纳米免疫传感器免疫响应电流与β-LG质量浓度的对数在2.5~100 ng/mL之间具有良好的线性关系,其检出限为0.75 ng/mL,同时该纳米免疫传感器具有良好的稳定性、特异性与重现性。该纳米免疫传感器用于实际样品β-乳球蛋白的检测,其加标回收率位于88.59%~97.64%之间,回收率较好,检测结果比ELISA精度更高,因此该法可用于乳制品中β-乳球蛋白过敏原的快速、精确检测。
[Key word]
[Abstract]
In this study, a nano-immunosensor was constructed through using chitosan, graphene and gold nanoparticles as the composite modification material to immobilize β-lactoglobulin (β-LG) antibody, to establish a rapid method for the determination of β-LG. The nano-immunosensor was characterized by cyclic voltammetry, using 1.0 mmol/L K3[Fe(CN)6] as the probe, and the experimental conditions were optimized. The optimal conditions were as follows: amount of the composite modification solution, 5 μL; pH of electrolyte solution, 7.0; loading amount of antibody, 40 ng; incubation temperature, 37 ℃; incubation time, 30 min. There was a good linear relationship between the immune-response current of the nano-immunosensor and the logarithm of β-lactoglobulin mass concentration at 2.5~100 ng/mL, with a limit of detection of 0.75 ng/mL. The nano-immunosensor exhibited good stability, specificity and reproducibility, and had a recovery rate of the spiked standard between 88.59% to 97.64% when it was applied to the determination of lactoglobulin in dairy products. The recovery rate was good, and the results were more accurate than those obtained by ELISA. Therefore, this methods can be used for rapid and accurate determination of β-lactoglobulin allergen in dairy products.
[中图分类号]
[基金项目]
河北省重点研发计划项目(17227113D)