基于原子转移自由基聚合反应的高灵敏度ELISA方法研究

摘要

Objective:To improve the traditional enzyme-linked immunosorbent assay(ELISA) strategy for increasing its sensitivity and extending the application.Methods:A large amount of horseradish peroxidase(HRP) was introduced to secondary antibody by introducing atom transfer radical polymer(ATRP) reaction and gold nanoparticles (GNPs).The modification improved the binding ratio of HRP to secondary antibody,achieved the object of detection signal amplification,and improved the traditional detection sensitivity greatly.Results:Limit of detection (LOD) after improvement decreased to 1/81 of that in traditional ELISA method.Conclusion:The newly built ELISA method had good stability,excellent sensitivity,and was able to detect ultralow-abundancy analytes.%目的:对传统酶联免疫吸附测定法(ELISA)进行优化改进,以提高其检测灵敏度,拓展应用范围.方法:通过在传统ELISA方法中引入原子转移自由基聚合反应(ATRP)和纳米金颗粒(GNPs),并将大量辣根过氧化物酶(HRP)与二抗结合,提高了HRP与抗原的结合比例,进而实现了对待检测物的信号放大.结果:优化后的ELISA方法在对β淀粉样蛋白的检测中,检测限(LOD)降低为原来的1/81.结论:改进后的ELISA方法性能稳定,灵敏度大幅提高,能够用于对痕量目标检测物的检测.