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Electrochemiluminescent aptamer biosensor for the determination of ochratoxin A at a gold-nanoparticles-modified gold electrode using N-(aminobutyl)-N-ethylisoluminol as a luminescent label

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Abstract

A highly selective electrochemiluminescent biosensor for the detection of target nephrotoxic toxin, ochratoxin A (OTA), was developed using a DNA aptamer as the recognition element and N-(4-aminobutyl)-N-ethylisoluminol (ABEI) as the signal-producing compound. The electrochemiluminescent aptamer biosensor was fabricated by immobilizing aptamer complementary DNA 1 sequence onto the surface of a gold-nanoparticle (AuNP)-modified gold electrode. ABEI-labeled aptamer DNA 2 sequence hybridized to DNA 1 and was utilized as an electrochemiluminescent probe. A decreased electrochemiluminescence (ECL) signal was generated upon aptamer recognition of the target OTA, which induced the dissociation of DNA 2 (ABEI-labeled aptamer electrochemiluminescent probe) from DNA 1 and moved it far away from the electrode surface. Under the optimal conditions, the decreased ECL intensity was proportional to an OTA concentration ranging from 0.02 to 3.0 ng mL-1, with a detection limit of 0.007 ng mL-1. The relative standard deviation was 3.8% at 0.2 ng mL-1 (n = 7). The proposed method has been applied to measure OTA in naturally contaminated wheat samples and validated by an official method. This work demonstrates the combination of a highly binding aptamer with a highly sensitive ECL technique to design an electrochemiluminescent biosensor, which is a very promising approach for the determination of small-molecule toxins.

In the absence of target, the ABEI-labeled aptamer probe is thought to hybridize with DNA 1 and assembled onto the surface of gold electrode, resulting in a strong ECL signal due to the tag close to the electrode surface. In the presence of target, the ABEI-labeled aptamer probe binds to OTA and dissociated from the surface of electrode, thus a small ECL signal generated attributed to the decrease of the ABEI-labeled aptamer probe assembled onto the electrode surface.

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Acknowledgements

This work was partly supported by the “863” project (2008AA10Z419), NSFC (20805019), High School Doctoral Programmes Founding of Ministry of Education (20070295014), NSF of Jiangsu Province (BK20081603), 111 project-B07029, and the Scientific and Technical Development Project of Qingdao in China (09-1-3-45-jch).

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Authors and Affiliations

  1. State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi, 214122, China

    Zhouping Wang, Nuo Duan & Shijia Wu

  2. MOE Key Laboratory of Eco-chemical Engineering; College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao, 266042, China

    Xu Hun

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  1. Zhouping Wang
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  2. Nuo Duan
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  3. Xu Hun
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  4. Shijia Wu
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Correspondence to Zhouping Wang.

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Wang, Z., Duan, N., Hun, X. et al. Electrochemiluminescent aptamer biosensor for the determination of ochratoxin A at a gold-nanoparticles-modified gold electrode using N-(aminobutyl)-N-ethylisoluminol as a luminescent label. Anal Bioanal Chem 398, 2125–2132 (2010). https://doi.org/10.1007/s00216-010-4146-1

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  • DOI: https://doi.org/10.1007/s00216-010-4146-1

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