Electrochemiluminescence Distance and Reactivity of Coreactants Determine the Sensitivity of Bead-Based Immunoassays |
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| Authors: | Dr Yafeng Wang Jialian Ding Dr Ping Zhou Jilin Liu Zhiyuan Qiao Kai Yu Prof Dr Jie Jiang Prof Dr Bin Su |
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| Institution: | 1. Key Laboratory of Excited-State Materials of Zhejiang Province, Department of Chemistry, Zhejiang University, Hangzhou, 310058 China;2. School of Chemistry and Chemical Engineering, Harbin Institute of Technology, Harbin, 150001 China;3. School of Environment, School of Marine Science and Technology (Weihai), Harbin Institute of Technology, Weihai, 150090 China |
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| Abstract: | Herein we report the study of electrochemiluminescence (ECL) generation by tris(2,2′-bipyridyl)ruthenium (Ru(bpy)32+) and five tertiary amine coreactants. The ECL distance and lifetime of coreactant radical cations were measured by ECL self-interference spectroscopy. And the reactivity of coreactants was quantitatively evaluated in terms of integrated ECL intensity. By statistical analysis of ECL images of single Ru(bpy)32+-labeled microbeads, we propose that ECL distance and reactivity of coreactant codetermine the emission intensity and thus the sensitivity of immunoassay. 2,2-bis(hydroxymethyl)-2,2′,2′′-nitrilotriethanol (BIS-TRIS) can well balance ECL distance-reactivity trade-off and enhance the sensitivity by 236 % compared with tri-n-propylamine (TPrA) in the bead-based immunoassay of carcinoembryonic antigen. The study brings an insightful understanding of ECL generation in bead-based immunoassay and a way of maximizing the analytical sensitivity from the aspect of coreactant. |
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| Keywords: | Electrochemiluminescence Immunoassay Radical Lifetime Reaction Mechanism Reactivity |
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