Study Of The Enhancement Of The Electrochemiluminescence Analysis System Based On N-butyldiethanolamine

Electrochemiluminescence （ECL） analytical technique has been received great attentiondue to its versatility, simplified optical setup, and good temporal and spatial control. The ECLof tris （2,2’-bipyridine） ruthenium（Ⅱ）/Tripropylamine is most widely studied and used. Withthe development of electrochemiluminescence analysis techniques and the need of setting upan immune detection method with ECL, building a new efficient electrochemiluminescencesystem is of great significance. The mechanisms of the ECL of tris （2,2’-bipyridine）ruthenium（Ⅱ）/Tripropylamine was briefly introduced firstly, the effective methods such asoptimizing the structure of the co-reactant，utilizing the surfactant were used for enhancingthe Ru（bpy）₃²⁺ECL intensity. In order to verify its superiority and feasibility, the newimmune analysis system were applied to detect influenza virus B（Flu B）.In order to understand the research background of this paper, theelectrochemiluminescence （ECL） and the methods for enhancing the Ru（bpy）₃²⁺ECLintensity was introduced simply. The detection methods of influenza virus B were alsodiscussed briefly.In order to improve the electrochemiluminescence （ECL） of Ru（bpy）₃²⁺, we report ancomparison of the efficiency of three common effective co-reactants such asN-butyldiethanolamine,2-（dibutylamino）ethanol and triethylamine at low Ru（bpy）₃²⁺concentration. N-butyldiethanolamine has been found to be the best co-reactant and showstrong ECL efficiency, then the optimal detection conditions of the Ru（bpy）₃²⁺/BDEA systemis studied. Under optimal conditions such as detection potential at1.8V, sample injection of150μL at30μL/s, the ECL intensity was linear with the concentrations of Ru（bpy）₃²⁺in therange of1×10^-9mol/L⁵×10^-7mol/L with a detection limit（S/N=3） of5×10^-10mol/L. Therelative standard deviations （RSD）<5%.In order to verify the feasibility and advantages of the newly established system, anelectroChemiluminescence immunoassay （ECLIA） for the detection of Flu B was established.The Flu B monoclonal antibody（mAb） was biotinylated，and the paired Ab was labeled withruthenium and the streptavidin-coated microparticles was used in this study．The ECLintensity was linear with the concentrations of antigen of Flu B in the range of0.55μg/ml and17.5μg/ml, with a detection limit of0.55μg/ml（S/N=3）. The precision，sensitivity andspecificity were evaluated．The established ECLIA for Flu B antigen in this study wasspecific，sensitive and precise. It may apply a convenient and rapid technique for clinical detection of Flu B.