Original Article
Nanostructured electrochemical biosensor for th0065 detection of the weak binding between the dengue virus and the CLEC5A receptor

https://doi.org/10.1016/j.nano.2014.03.009Get rights and content

Abstract

In this paper, we develop an effective method for detecting weak molecular bonding between the dengue virus (DV) and its receptor C-type lectin domain family 5, member A (CLEC5A). The CLEC5A–DV interaction is critical for DV-induced hemorrhagic fever and shock syndrome, so the sensing of CLEC5A–DV binding is crucial to realize a thorough study of the pathogenesis of dengue fever. Through a highly sensitive nanostructured sensing electrode of gold nanoparticles (GNPs) uniformly deposited on a nanohemisphere array, a label-free detection of the ultra weak binding between CLEC5A and the DV can be performed with electrochemical impedance spectroscopy (EIS). Experimental results demonstrate that the proposed approach is a highly promising method for investigating weak molecular interactions such as the ligand–receptor interaction of dengue fever, enterovirus (EV), or the interaction between cancer surface glycoproteins and their receptors.

From the Clinical Editor

Authors of this study investigated the ultra-weak binding between dengue virus and its CLEC5A receptor via electrochemical impedance spectroscopy and gold NP sensing electrode. Similar methods may be applicable in other infections and in cancer models as well.

Graphical Abstract

An effective method for detecting weak molecular bonding between the dengue virus (DV) and its receptor C-type lectin domain family 5, member A (CLEC5A) has been developed. Label-free detections of the ultra weak binding were carried out via a highly sensitive nanostructured sensing electrode with gold nanoparticles (GNPs) uniformly deposited on a nanohemisphere array. Experimental results demonstrate that the proposed approach is a highly promising method for investigating weak molecular interactions such as the ligand–receptor interaction of DV and enterovirus.

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Section snippets

Fabrication of nanostructured biosensor

The composition of the nanostructured sensor chip is shown in Figure 1. To increase the surface reacting area, the barrier-layer surface of an AAO membrane was used as the substrate of the sensor chip. The AAO membrane was prepared by the conventional anodization process, which was described in our previous study.15 After anodic oxidization, the non-oxidized aluminum beneath the barrier layer was removed by CuCl2 · HCl solution, which was prepared by dissolving 13.45 g of CuCl2 powder in 100 ml of

Characterization of nanostructured sensing surface

The nanostructure of our sensor surface was analyzed using a scanning electron microscope (SEM) (Figure 4). Through our fabrication process, a modified AAO barrier layer of uniformly distributed nano-hemispheres with a diameter about 200 nm was formed (Figure 4, A). GNPs with an average diameter of less than 10 nm were uniformly and compactly deposited on the orderly hemispheric electrode array (Figure 4, B). The uniformly scattering electric flux that is perpendicular to the nanohemispheric

Discussion

The common approach used to realize high-throughput screening in protein–protein interaction is by using the ELISA or surface plasmon resonance (SPR) technique. ELISA has the advantages of low cost and simple operation; however, its relatively low sensitivity limits the detection of weak molecular-bonding such as the CLEC5A–DV interactions. The SPR technique is a high-sensitivity method used to screen protein–protein interactions, which has the added advantage of being label-free. However, its

References (29)

  • N. Gunasekara et al.

    Characterization of a novel weak interaction between MUC1 and Src-SH3 using nuclear magnetic resonance spectroscopy

    Biochem Biophys Res Commun

    (2012)
  • A.A. Watson et al.

    Structural flexibility of the macrophage dengue virus receptor CLEC5A: implications for ligand binding and signaling

    J Biol Chem

    (2011)
  • N.D. Rambaruth et al.

    Cell surface glycan–lectin interactions in tumor metastasis

    Acta Histochem

    (2011)
  • A.T. Back et al.

    Dengue viruses — an overview

    Infect Ecol Epidemiol

    (2013)
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    Role of the funding source: This publication was supported by the Department of Health of Taiwan (DOH102-TD-N-111-004), National Science Council of Taiwan (NSC-101-2212-E-005-022- MY3), National Science Council (NSC 101-2325-B-010-006 and NSC 101-2321-B-010-003), Academia Sinica, and Thematic Research Project (AS-101-TP-B06-2), and in part by the Infection and Immunity Center, National Yang-Ming University, Taiwan (grant from Ministry of Education, Aim for the Top University Plan), Taipei Veterans General Hospital (V101E4-006, V101E4-007 and TVGH-NTUH VN-100-06), and the Molecular and Genetic Imaging Core/National Research Program for Genomic Medicine at National Yang-Ming University (NSC99-3112-B-010-015).

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