A sensitive and rapid immunoassay for quantification of testosterone by microchip electrophoresis with enhanced chemiluminescence detection
2011
Huang, Yong | Shi, Ming | Zhao, Shulin | Liang, Hong
A sensitive and rapid approach to perform testosterone (T) competitive immunoassay by microchip electrophoresis (MCE) with chemiluminescence (CL) detection was developed. The assay is based on the competitive immunoreactions between T and Nâ(4âaminobutyl)âNâethylisoluminolâlabeled T (ABEIâT) with a limited amount of antibody (Ab), and the rapid electrophoretic separation of an equilibrated mixture of ABEIâTâAb complex and free ABEIâT, followed by CL detection using horseradish peroxidaseâcatalyzed ABEIâH2O2 system. Free ABEIâT and the ABEIâTâAb complex are well separated within 30âs under the assay conditions. The developed method could be used to determine T with good precision and a detection limit lower than 1.0ânM. This method was applied for the quantification of T in human serum. The results demonstrated that the current MCEâCLâbased competitive immunoassay maybe served as an alternative tool for clinical analysis.
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