title: Development of double-generation gold nanoparticle chip-based dengue virus detection system combining fluorescence turn-on probes.
abstract: A sensing platform, combined amino acid labeling kit and a double-generation gold nanoparticle (DG-AuNP) chip, was designed to prove the existence of weak but crucial binding between the DV (dengue virus) and its CLEC5A receptor. At first, we have designed a kit combining the novel fluorescence turn-on sensors for lysine, arginine and cysteine amino acids. Advantages of this kit are that emission on-off switching can increase the signal-to-noise ratio and the virus must be fluorescently labelled with sufficient numbers of fluorophores because the lysine, arginine and cysteine residues of viral proteins are labelled simultaneously. Consequently, this kit can be used to light-on single DV spot both in solution and in cell under fluorescence microscopy. Second, the labeling kit was used to examine the DV binding to the CLEC5A-coated DG-AuNP chip. Based on our study, the double-generation gold nanoparticle construction of chip can support more coating areas for receptor CLEC5A and then, support more binding opportunities for DV. Meanwhile, the grooves between nanohemispheres will provide the extra driving force for DV stacking. We try to give a proof that this sensing platform is very useful for detection of weak binding mechanism.
authors.affiliation: Ph.D. Program in Tissue Engineering and Regenerative Medicine, National Chung-Hsing University, Taichung, Taiwan.
authors.forename: Yen-Ting
authors.initials: YT
authors.lastname: Tung
authors.affiliation: Graduate Institute of Biomedical Engineering, National Chung Hsing University, 250 Kuo Kuang Road, Taichung 402, Taiwan. Electronic address: ccchang555@dragon.nchu.edu.tw.
authors.forename: Cheng-Chung
authors.initials: CC
authors.lastname: Chang
authors.affiliation: Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan.
authors.forename: Yi-Ling
authors.initials: YL
authors.lastname: Lin
authors.affiliation: Genomics Research Center, Academia Sinica, Taipei 115, Taiwan; Institute of Clinical Medicine, National Yang-Ming University, Taipei 112, Taiwan.
authors.forename: Shie-Liang
authors.initials: SL
authors.lastname: Hsieh
authors.affiliation: Ph.D. Program in Tissue Engineering and Regenerative Medicine, National Chung-Hsing University, Taichung, Taiwan; Graduate Institute of Biomedical Engineering, National Chung Hsing University, 250 Kuo Kuang Road, Taichung 402, Taiwan. Electronic address: gjwang@dragon.nchu.edu.tw.
authors.forename: Gou-Jen
authors.initials: GJ
authors.lastname: Wang
mesh_terms: D003716:Dengue Virus / Q000302:isolation & purification*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005456:Fluorescent Dyes / Q000737:chemistry; D006046:Gold / Q000737:chemistry*; D053768:Metal Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D008856:Microscopy, Fluorescence / Q000295:instrumentation*; D010316:Particle Size; D013050:Spectrometry, Fluorescence / Q000295:instrumentation*; D019562:Viral Load / Q000295:instrumentation*
chemical_list: D005456:Fluorescent Dyes; D006046:Gold
keywords: CLEC5A receptor; DV (dengue virus); Fluorescence microscopy; Fluorescent turn-on probes; Gold nanoparticle; Nanohemispheres