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Fabrication of Troponin I Biosensor Composed of Multi-Functional DNA Structure/Au Nanocrystal Using Electrochemical and Localized Surface Plasmon Resonance Dual-Detection Method

1
Department of Chemical Engineering, Kwangwoon University, Wolgye-dong, Nowon-gu, Seoul 01899, Korea
2
Department of Chemical Engineering and Materials Science, Chung-Ang University, Heukseok-dong, Dongjak-gu, Seoul 06974, Korea
3
Human IT Convergence Research Center, Korea Electronics Technology Institute, Seongnam-si, Gyeonggi-do 13509, Korea
4
Department of Chemical and Biomolecular Engineering, Sogang University, 35 Baekbeom-ro, Mapo-gu, Seoul 04107, Korea
5
School of Chemical and Biological Engineering, Institute of Chemical Process, Seoul National University, Seoul 08826, Korea
6
School of Integrative Engineering Chung-Ang University, Heukseok-dong, Dongjak-gu, Seoul 06974, Korea
*
Authors to whom correspondence should be addressed.
Nanomaterials 2019, 9(7), 1000; https://doi.org/10.3390/nano9071000
Received: 13 June 2019 / Revised: 30 June 2019 / Accepted: 5 July 2019 / Published: 11 July 2019
(This article belongs to the Special Issue Application of Nanotechnology in Cardiology)
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Abstract

In the present study, we fabricated a dual-mode cardiac troponin I (cTnI) biosensor comprised of multi-functional DNA (MF-DNA) on Au nanocrystal (AuNC) using an electrochemical method (EC) and a localized surface plasmon resonance (LSPR) method. To construct a cTnI bioprobe, a DNA 3 way-junction (3WJ) was prepared to introduce multi-functionality. Each DNA 3WJ arm was modified to possess a recognition region (Troponin I detection aptamer), an EC-LSPR signal generation region (methylene blue: MB), and an anchoring region (Thiol group), respectively. After an annealing step, the multi-functional DNA 3WJ was assembled, and its configuration was confirmed by Native-TBM PAGE for subsequent use in biosensor construction. cTnI was also expressed and purified for use in biosensor experiments. To construct an EC-LSPR dual-mode biosensor, AuNCs were prepared on an indium-tin-oxide (ITO) substrate using an electrodeposition method. The prepared multi-functional (MF)-DNA was then immobilized onto AuNCs by covalent bonding. Field emission scanning electron microscope (FE-SEM) and atomic force microscopy (AFM) were used to analyze the surface morphology. LSPR and electrochemical impedance spectroscopy (EIS) experiments were performed to confirm the binding between the target and the bioprobe. The results indicated that cTnI could be effectively detected in the buffer solution and in diluted-human serum. Based on the results of these experiments, the loss on drying (LOD) was determined to be 1.0 pM in HEPES solution and 1.0 pM in 10% diluted human serum. Additionally, the selectivity assay was successfully tested using a number of different proteins. Taken together, the results of our study indicate that the proposed dual-mode biosensor is applicable for use in field-ready cTnI diagnosis systems for emergency situations. View Full-Text
Keywords: dual-mode biosensor; cardiac troponin I biosensor; multi-functional DNA structure; electrochemical biosensor; LSPR; EC-LSPR dual-mode biosensor; cardiac troponin I biosensor; multi-functional DNA structure; electrochemical biosensor; LSPR; EC-LSPR
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

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Lee, T.; Kim, J.; Nam, I.; Lee, Y.; Kim, H.E.; Sohn, H.; Kim, S.-E.; Yoon, J.; Seo, S.W.; Lee, M.-H.; Park, C. Fabrication of Troponin I Biosensor Composed of Multi-Functional DNA Structure/Au Nanocrystal Using Electrochemical and Localized Surface Plasmon Resonance Dual-Detection Method. Nanomaterials 2019, 9, 1000.

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