Development of neuraminidase detection using gold nanoparticles boron-doped diamond electrodes

Significance Statement

Neuraminidase detection is substantial in controlling the disease caused by related virus and microbes since Neuraminidase is an important enzyme in the pathogenic viruses and microbes spreading. Common detection methods of neuraminidase, such as RT-PCR, ELISA, and enzymatic reaction require several types of chemical reagents, specific instruments, and highly skilled operators. Therefore, a more simple and practical method is urgently required. In this work, a sensor for Neuraminidase has been developed using gold nanoparticles modified boron-doped diamond (AuNPs-BDD) electrodes. The detection method was performed based on the difference of electrochemical responses of Zanamivir at gold surface before and after the reaction with neuraminidase in phosphate buffer solution (PBS) pH 5.5.

In order to have a stable response, the modified electrodes was prepared through a self-assembly deposition of AuNPs at an amine-terminated boron-doped diamond. Cyclic voltammetry was used for the method. A linear calibration curve for Zanamivir in 0.1 M PBS in the absence of neuraminidase was achieved in the concentration range of 1 x 10^-6 – 1 x 10^-5 M (R² = 0.99), with the estimated limit of detection (LOD) of 2.29 x 10^-6 M.

Furthermore, using its reaction with 1.00 x 10^-5 M Zanamivir a linear calibration curve of neuraminidase can be obtained in the concentration range of 0 – 12 mU (R² = 0.99) with an estimated LOD of 0.12 mU. High reproducibility was shown with an RSD of 1.14 % (n=30). These performances could be maintained when the detection was performed in mucin matrix. Comparison made using gold-modified BDD (Au-BDD) electrodes suggested that the good performance of the detection method is due to the stability of the gold particles position at the BDD surface.

About the author

Dr. Tribidasari A. Ivandini is a member of the teaching and research staff at Department of Chemistry, Faculty of Mathematics and Science, Universitas Indonesia, since 1997. She was born in Jakarta, Jakarta, on January 29th, 1970. She received her magister degrees in 1996 from Universitas Indonesia. Then, in 2003 she accomplished her doctoral degree at The University of Tokyo, Tokyo, Japan. Between 2003 and 2007 she performed post-doctoral research at the Department of Chemistry, Keio University, Tokyo, Japan with JSPS fellowship during 2004-2006. Up to the present time she is still performing research collaboration with Keio University.

Her research interests are mostly concerning electrochemistry of diamond, including its development for sensors and biosensors, electrocatalytics, and waste-water treatments. She has published about 50 scientific papers in highly respectable journals and conference proceedings, such as Analytical Chemistry, Physical Chemistry Chemical Physics, Sensors Actuators B: Chemicals, and Diamond Related Materials. She also holds some patents under European and Japan Patents. Ivandini can be reached at the Department of Chemistry, Faculty of Mathematics and Science, Universitas Indonesia, Indonesia.E-mail : [email protected]

About the author

Dr. Yasuaki Einaga is a professor in the Department of Chemistry at Keio University, Japan. He received his BS (1994) , MS (1996) , and PhD (1999) degree from The University of Tokyo. After 2 years as a research associate at The University of Tokyo, he started a faculty career as an assistant professor in Keio University in 2001. He was promoted to associate professor in 2003, and to professor in 2011. He was also a research director of JST-CREST (2011-2014), and JST-ACCEL (2014-present). His research interests include functional materials science, photochemistry, electrochemistry, and diamond electrodes. Einaga can be reached at the Department of Chemistry, Keio University, Yokohama, Japan. E-mail [email protected]

Journal Reference

Anal Biochem. 2016 Mar 15;497:68-75.

Wahyuni WT¹, Ivandini TA², Saepudin E¹, Einaga Y³.

[expand title=”Show Affiliations”]

Department of Chemistry, Faculty of Mathematics and Sciences, University of Indonesia, Kampus UI Depok, Jakarta 16424, Indonesia.
Department of Chemistry, Faculty of Mathematics and Sciences, University of Indonesia, Kampus UI Depok, Jakarta 16424, Indonesia. Electronic address: [email protected].
Department of Chemistry, Faculty of Science and Technology, Keio University, Yokohama 223-8522, Japan; JST ACCEL, Yokohama 223-8522, Japan. Electronic address: [email protected].

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Abstract

Gold nanoparticles-modified boron-doped diamond (AuNPs-BDD) electrodes, which were prepared with a self-assembly deposition of AuNPs at amine-terminated boron-doped diamond, were examined for voltammetric detection of neuraminidase (NA). The detection method was performed based on the difference of electrochemical responses of zanamivir at gold surface before and after the reaction with neuraminidase in phosphate buffer solution (PBS, pH 5.5). A linear calibration curve for zanamivir in 0.1 M PBS in the absence of NA was achieved in the concentration range of 1 × 10(-6) to 1 × 10(-5) M (R(2) = 0.99) with an estimated limit of detection (LOD) of 2.29 × 10(-6) M. Furthermore, using its reaction with 1.00 × 10(-5) M zanamivir, a linear calibration curve of neuraminidase can be obtained in the concentration range of 0-12mU (R(2) = 0.99) with an estimated LOD of 0.12mU. High reproducibility was shown with a relative standard deviation (RSD) of 1.14% (n = 30). These performances could be maintained when the detection was performed in mucin matrix. Comparison performed using gold-modified BDD (Au-BDD) electrodes suggested that the good performance of the detection method is due to the stability of the gold particles position at the BDD surface.

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