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Electrochemical sandwich immunoassay for insulin detection based on the use of gold nanoparticle-modified MoS2 nanosheets and the hybridization chain reaction

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Abstract

A sandwich-type of electrochemical immunoassay is described for the determination of insulin. It is based on the use of a glassy carbon electrode that was modified with MoS2 nanosheets decorated with gold nanoparticles (AuNPs) to immobilize a large amount of first antibody (Ab1). Following exposure to insulin, secondary antibody (Ab2) that was cross-linked to a DNA initiator strand (T0) to form an Ab2@T0 conjugate was added to undergo a sandwich immunoreaction. Subsequently, the long dsDNA concatemer was formed by a hybridization chain reaction between Ab2@T0 and auxiliary probes (H1, H2). Finally, the electrochemical probe ruthenium(II) hexaammine was intercalated into the dsHCR products via electrostatic interaction between the anionic DNA phosphate backbones and the cationic probe. The electrochemical response, best measured at a potential of around −0.21 V (vs Ag/AgCl) has a dynamic range that extends from 0.1 pmol L−1 to 1 nmol L−1 insulin, and the detection limit is as low as 50 fmol L−1. The assay was acceptably specific, reproducible and stable. In our perception, it represents a viable new tool for determination of this important clinical parameter.

Schematic of a sandwich-type of electrochemical immunoassay for the determination of insulin based on the use of MoS2 nanosheets modified with gold nanoparticles (AuNP@MoS2) and hybridization chain reaction (HCR).

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Scheme 1
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Abbreviations

Ab1 :

First antibody

Ab2 :

Secondary antibody

BSA:

Bovine serum albumin

[Ru(NH3)6]3+ :

Hexaammineruthenium(III) dichloride

H1 and H2 :

Auxiliary probes

AuNP@MoS2 :

Molybdenum disulfide nanosheets modified with gold nanoparticles

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Acknowledgments

This work was funded by the National Natural Science Foundation of China (81572080), and the Special Project for Social Livelihood and Technological Innovation of Chongqing (cstc2015shmszx120040 and cstc2016shmszx130043).

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Author notes

  1. Huidan Sun and Shaoyan Wu contributed equally to this work.

Authors and Affiliations

  1. Medical Examination Centre, the First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China

    Huidan Sun, Shaoyan Wu, Rong Luo & Shijia Ding

  2. Department of Endocrinology, the First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China

    Huidan Sun & Rong Luo

  3. Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University, Chongqing, 400016, China

    Xiaoyan Zhou, Min Zhao, Haiping Wu & Shijia Ding

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  1. Huidan Sun
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  2. Shaoyan Wu
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  4. Min Zhao
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Correspondence to Rong Luo or Shijia Ding.

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Sun, H., Wu, S., Zhou, X. et al. Electrochemical sandwich immunoassay for insulin detection based on the use of gold nanoparticle-modified MoS2 nanosheets and the hybridization chain reaction. Microchim Acta 186, 6 (2019). https://doi.org/10.1007/s00604-018-3124-8

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  • DOI: https://doi.org/10.1007/s00604-018-3124-8

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