改进酶联免疫吸附新方法检测甲胎蛋白
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摘要:
将纳米金(GNP,gold nanoparticle)功能化修饰的一抗(Ab1,first antibody)和氧化石墨烯(GO,graphene oxide)功能化修饰的二抗(Ab2,second antibody)通过原子转移自由基聚合反应(ATRP,atom transfer radical polymerization)应用于酶联免疫吸附(ELISA,enzyme linked immunosorbent assay)方法,提高检测灵敏度.考察了纳米材料的信号逐级放大效果,得出最佳条件为:辅助一抗/一抗(aAb1/Ab1,accessory antibody/Ab1)比值为3∶2,辣根过氧化物酶/二抗(HRP/Ab2,horseradish peroxidase/Ab2)比值为160∶1,在此条件下对甲胎蛋白(AFP,alpha fetoprotein)进行检测.结果表明,改进后的GNP/ATRP/GO-ELISA(GAG-ELISA)方法灵敏度提高了729倍,而成本仅为传统ELISA法的1/20.该研究提出的将多种纳米材料同时应用到传统检测方法中,实现信号逐级放大的思路是可行的,也为其他检测方法的改进提供了借鉴.
In this paper,Gold nano particle(GNP)functionalized first antibody(Ab1)and graphene oxide(GO)functionalized second antibody(Ab2)were applied to enzyme-linked immunosorbent assay(ELISA)by atom transfer radical polymerization(ATRP)to improve the detection sensitivity.The results show that the ratio of aAb1/Ab1 and HRP/Ab2 is 3∶2 and 160∶1 respectively.Under these conditions,AFP was detected.The results showed that the sensitivity of the improved GNP/ATRP/GO-ELISA(GAG-ELISA)was increased by 729 times,and the cost was only one twentieth of that of the traditional ELISA.In this research,the idea of applying a variety of nano materials to the traditional detection method at the same time to realize the step-by-step signal amplification is feasible,and also provides a reference for other detection methods.
作者:
漆璐 林虹君 张爱红 王兴河
Qi Lu;Lin Hongjun;Zhang Aihong;Wang Xinghe(Phase I Clinical Trial Center,Beijing Shijitan Hospital,Capital Medical University,Beijing 100038,China;Unit 32311 of PLA,Beijing 102112,China;Institute of Chemical Defense of PLA,Beijing 102205,China)
机构地区:
首都医科大学附属北京世纪坛医院药物Ⅰ期临床试验研究室 中国人民解放军32311部队 中国人民解放军陆军防化学院
出处:
《betway官方app 学报：自然科学版》 CAS 北大核心 2020年第5期87-92,共6页
基金:
国家科技重大专项-重大新药创制基金资助项目(2017ZX09304026) 北京市科委关键技术平台“靶向性创新药物人体零期研究平台建设”(20180808).
关键词:
酶联免疫吸附 氧化石墨烯 原子自由基聚合 纳米金
enzyme linked immunosorbent assay graphene oxide atom transfer radical polymerization nano gold
分类号:
Q533 [生物学—生物化学]
