Original ArticleNanostructured electrochemical biosensor for th0065 detection of the weak binding between the dengue virus and the CLEC5A receptor
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.
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
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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).