Ultrasensitive electrochemiluminescence immunosensor based on luminol functionalized gold nanoparticle labeling

电化学发光 鲁米诺 检出限 分析物 化学 链霉亲和素 胶体金 共轭体系 化学发光 免疫分析 色谱法 纳米颗粒 结合 纳米技术 组合化学 生物素 材料科学 聚合物 抗体 生物化学 有机化学 数学分析 生物 免疫学 数学
作者
Dayong Tian,Chunfeng Duan,Wei Wang,Hua Cui
出处
期刊:Biosensors and Bioelectronics [Elsevier BV]
卷期号:25 (10): 2290-2295 被引量:123
标识
DOI:10.1016/j.bios.2010.03.014
摘要

An ultrasensitive electrochemiluminescence (ECL) immunosensor based on luminol functionalized gold nanoparticle (AuNP) labeling was developed using human immunoglobulin G (hIgG) as a model analyte. The primary antibody biotin-conjugated goat-anti-human IgG was first immobilized on a streptavidin coated AuNP modified electrode, then the antigen (human IgG) and the luminol functionalized AuNP-labeled second antibody were conjugated successively to form a sandwich-type immunocomplex, i.e. immunosensor. ECL was carried out with a double-step potential in carbonate buffer solution containing 1.0 mmol/L H(2)O(2). Since thousand of luminol molecules were coated on the surface of AuNPs to realize labeling of multiple molecules with CL activity at a single antibody and the amplification of AuNPs and biotin-streptavidin system was utilized, luminol ECL signal could be enhanced greatly, finally resulting in extremely high sensitivity. The ECL method shows a detection limit of 1.0 pg/mL (S/N=3) for hIgG, which is superior to all previously reported methods for the determination of hIgG. Moreover, the proposed method is also simple, stable, specific, and time-saving, avoiding the complicated stripping procedure during CL detection and the uncontrollable synthesis of irregular nanoparticles compared with other chemiluminescence immunoassay based on AuNP labeling. Additionally, the labeling procedure is also superior to that of other reported multilabeling strategies, such as Ru complex-encapsulated polymer microspheres, and most of Ru complex-encapsulated liposomes in simplicity, stability, labeling property and practical applicability. Finally, the proposed method has been successfully applied to the detection of hIgG in human serums.
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