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静电作用力引发双链 DNA 的弹性凝聚用于高性能一步法免疫分析。

Electrostatic Force Triggering Elastic Condensation of Double-Stranded DNA for High-Performance One-Step Immunoassay.

机构信息

College of Chemistry and Chemical Engineering , Central South University , Changsha 410083 , P. R. China.

State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering , Hunan University , Changsha 410082 , P. R. China.

出版信息

Anal Chem. 2018 Oct 2;90(19):11446-11452. doi: 10.1021/acs.analchem.8b02556. Epub 2018 Sep 13.

Abstract

Current strategies for high-performance immunoassay generally require a sandwich structure for signal amplification. This strategy is limited to multivalent antigens and complicates the detection scheme. Herein we demonstrate a class of simple one-step ultrasensitive immunoassay with the adoption of double-stranded DNA (dsDNA) as "conductive spring" to bridge the electrode and redox-reporter/antibody-receptor comodified gold nanoparticles (AbFc@AuNPs). Upon biorecognition between antigen and antibody, the charge of the AuNPs changes, enhancing the electrostatic interaction between the AuNPs and Au electrode surface, and condensing the dsDNA chain. For the first time, the sensitive response of the electrochemical redox current to the DNA chain length is utilized to achieve an ultrahigh sensitivity down to fM level. Only the primary antibody needed in the recognition interface ensures the one-step immunoreaction works well with monovalent antigens, which ensure this method as a promising general alternative means for fast, high-throughput or point-of-care clinical applications even for very challenging clinically relevant samples.

摘要

目前用于高性能免疫分析的策略通常需要采用三明治结构进行信号放大。这种策略仅限于多价抗原,并且使检测方案复杂化。在此,我们采用双链 DNA(dsDNA)作为“导电弹簧”来桥接电极和氧化还原报告物/抗体受体共修饰的金纳米颗粒(AbFc@AuNPs),展示了一类简单的一步式超灵敏免疫分析。在抗原和抗体之间发生生物识别后,AuNPs 的电荷发生变化,增强了 AuNPs 和 Au 电极表面之间的静电相互作用,并使 dsDNA 链凝结。首次利用电化学氧化还原电流对 DNA 链长的敏感响应来实现超灵敏检测,灵敏度可低至 fM 级。仅在识别界面中需要的主要抗体即可确保一步免疫反应能够很好地与单价抗原结合,这确保了该方法成为一种有前途的通用替代手段,可用于快速、高通量或即时护理临床应用,即使是非常具有挑战性的临床相关样本也是如此。

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