基于多重信号放大的夹心式甲胎蛋白免疫传感器
sandwich-type AFP immunosensor based on multiple signal amplification
本文制备了新型三维有序多孔(3DOM)聚多巴胺-纳米金(PDA-AuNPs)修饰电极作为传感界面、核壳型磁性纳米复合物CoFe2O4@SiO2/自掺杂聚苯胺纳米纤维(SPANF)-金纳米粒子(AuNPs)作为二抗标记的纳米探针,构建了夹心式甲胎蛋白(AFP)电化学免疫传感器。以SiO2胶体晶体为模板制备了3DOM PDA仿生膜,原位修饰了AuNPs后,形成了导电性好、生物相容性好的3DOM PDA-AuNPs仿生界面,用来固定AFP 一抗(Ab1)形成传感识别界面。CoFe2O4@SiO2/SPANF-AuNPs复合材料具备了磁性、生物相容性、导电性等重要性质用于磁性分离及固定二抗。当溶液中AFP分子被Ab1识别后,标记有CoFe2O4@SiO2/SPANF-AuNPs的二抗(Ab2)立即被AFP捕获。以SPAN作为信号来源,在最佳实验条件下,对AFP响应的线性范围为0.05-10.05 ng mL-1,检测限为0.005 ng mL-1。另外,将该传感器用于实际血清样品的检测,结果与临床电化学免疫发光法测定结果一致,展示了该传感器的应用前景。
In this paper, a novel sandwich-type immunosensor protocol for the detection of alpha-fetoprotein (AFP) was designed using a three dimensional ordered macroporous polydopamine-Au nanoparticles hybird (3DOM PDA-AuNPs) biofilm as platform, and core-shell structured magnetic nanocomposites of CoFe2O4@SiO2/self-doped polyaniline nanofiber (SPANF)-AuNPs as nano probe. The 3DOM PDA biomimic membrane was fabricated by electrodeposition of dopamine (DA) within the interstices of SiO2 colloidal template. After in situ deposition of AuNPs on PDA film, the 3DOM PDA-AuNPs with high conductivity and fantastic biocompatibility was formed to immobilize primary antibody (Ab1). The CoFe2O4@SiO2/SPANF-AuNPs nanocomposite can be used for magnetic separation and the immobilization of the secondary antibody (Ab2) due to the properties of magnetism and biocompatibility. When AFP in the solution was recognized by Ab1, the CoFe2O4@SiO2/SPANF-AuNPs labeled Ab2 could be immediately combined with AFP. Under the optimal conditions and using SPAN as the signal source, the immunosensor exhibited a good linear response in the range of 0.05 to 10.05 ng mL-1, with the detection limit of 0.005 ng mL-1. Moreover, the fabricated immunosensor was also evaluated for real serum sample, obtaining an acceptable agreement with the results from the commercialized electrochemiluminescent (ECL) automatic analyzer. In this regard, this new type of amplification strategy is proved to be a promising and reliable tool for rapid detection of AFP and may open up a wide horizon for the clinic application as well.
钱春花、陈晓君、许小龙
生物科学研究方法、生物科学研究技术生物工程学基础医学
三维有序大孔材料聚多巴胺-纳米金磁性核壳复合材料夹心式免疫传感器甲胎蛋白
hree dimensional ordered macroporous nanomaterialPolydopamine-Au nanoparticles hybridCore-shell structured magnetic nanocompositesSandwich-type electrochemical immunosensorAlpha-fetoprotein.
钱春花,陈晓君,许小龙.基于多重信号放大的夹心式甲胎蛋白免疫传感器[EB/OL].(2015-04-14)[2025-08-18].http://www.paper.edu.cn/releasepaper/content/201504-221.点此复制
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