Neutravidin biosensor for direct capture of dual-functional biotin-molecular beacon-AuNP probe for sensitive voltammetric detection of microRNA

SENSORS AND ACTUATORS B-CHEMICAL

卷:248

Article

Fredj, Zina^1,2  Azzouzi, Sawsen^1,2  Turner, Anthony P. F.¹  Ben Ali, Mounir²  Mak, Wing Cheung¹ 

[1] Linkoping Univ, Dept Phys Chem & Biol IFM, Biosensors & Bioelect Ctr, S-58183 Linkoping, Sweden

[2] Univ Sousse, Higher Inst Appl Sci & Technol Sousse, ISSAT, GREENS, Cite Ettafala 4003, Ibn Khaldoun Sousse, Tunisia

关键词: Neutravidin;    Biosensor;    microRNA;    Molecular beacon;    Stripping voltammetry;   

DOI  :  10.1016/j.snb.2017.03.160

来源: Elsevier

【 摘 要 】

We have demonstrated a new approach using a neutravidin-based biosensor combined with a dual-function gold nanoparticle (AuNP) biolabel, for simple and sensitive detection of microRNA-21 (miRNA-21). The selectivity of the biosensor is provided by the intrinsic properties of the dual-functional biotin-MB-AuNP label. The assay procedure is relatively simple, exploiting a one-pot assay concept where the affinity capture of the miRNA-21/dual-functional biotin-MB-AuNP complex, via the strong biotinneutravidin supramolecular interaction, and simultaneous detection of the captured AuNP5 label with stripping voltammetry, is performed in a single step. This electrochemical miRNA biosensor could detect miRNA-21 with limit of detection of 0.1 x 10(-12) and a dynamic range from 0.5 x 10(-1)2 to 1.0 x 10(-9)M. The performance of the miRNA-21biosensor was further improved after silver deposition onto the AuNP5, delivering an enhanced detection limit of 4.0 x 10(-15)M of miRNA-21, and an extremely wide analytic dynamic range from 10 x 10(-15) to 1 x 10(-9)M (5 orders of magnitude). This exceptionally broad dynamic range demonstrates the advantage of the one-pot assay approach with direct capture of the dual functional biotin-MB-AuNP via the strong biotin-neutravidin supramolecular interaction. Furthermore, we demonstrated the detection of miRNA-21 in spiked serum at clinically relevant concentrations. The miRNA biosensor displayed excellent analytical performance for the detection of miRNA and could provide a powerful and convenient tool for biomedical research and applications in cancer diagnostics. (C) 2017 Elsevier B.V. All rights reserved.

【 授权许可】

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