An Immunosensing System Using Stilbene Glycoside as a Fluorogenic Substrate for an Enzymatic Reaction ModelReportar como inadecuado




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College of Chemistry and Biological Engineering, Changsha University of Science and Technology, Changsha, 410076, Hunan, P.R. China





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Abstract A natural product, stilbene glycoside 2,3,5,4’-tetrahydroxydiphenylethylene-2-O-glucoside, TBG, has been evaluated for the first time as a potential substrate for horseradish peroxidase HRP-catalyzed fluorogenic reactions. The properties of TBG as a fluorogenic substrate for HRP and its application in a fluorometric enzyme-linked immunosensing system were compared with commercially available substrates such as p-hydroxyphenylpropionic acid pHPPA, chavicol and Amplex red using Brucella melitensis antibody BrAb as a model analyte. The immunosensing body based on HRP-BrAb was constructed by dispersing graphite, BrAg and paraffin wax at room temperature. In a competitive immunoassay procedure, the BrAb competed with HRP-BrAb to react with the immobilized BrAg. In the enzymatic reaction, the binding HRP-BrAb on the sensing body surface can catalyze the polymerization reaction of TBG by H2O2 forming fluorescent dimers and causing an increase in fluorescence intensity. TBG showed comparable ability for HRP detection and its enzyme-linked immunosensing reaction system, in a linear detection ranging of 3.510-8~7.610-6g-L and with a detection limit of 1.710-9 g-L. The immobilized biocomposite surface could be regenerated with excellent reproducibility RSD=3.8% by simply polishing with an alumina paper. The proposed immunosensing system has been used to determine the BrAb in rabbit serum samples with satisfactory results. View Full-Text

Keywords: Stilbene glycoside; HRP fluorogenic substrate; enzyme-linked immunosensing; Brucella melitensis Stilbene glycoside; HRP fluorogenic substrate; enzyme-linked immunosensing; Brucella melitensis





Autor: Xue-Hui Zhan, Fu-Chun Gong * , Shu-Zhen Tan, Peng-Mian Huang and Ya-Fei Tan

Fuente: http://mdpi.com/



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