A sensitive and rapid immunoassay for quantification of testosterone by microchip electrophoresis with enhanced chemiluminescence detection.
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 N-(4-aminobutyl)-N-ethylisoluminol-labeled T (ABEI-T) with a limited amount of antibody (Ab), and the rapid electrophoretic separation of an equilibrated mixture of ABEI-T-Ab complex and free ABEI-T, followed by CL detection using horseradish peroxidase-catalyzed ABEI-H(2)O(2) system. Free ABEI-T and the ABEI-T-Ab complex are well separated within 30  s under the assay conditions. The developed method could be used to determine T with good precision and a detection limit lower than 1.0  nM. This method was applied for the quantification of T in human serum. The results demonstrated that the current MCE-CL-based competitive immunoassay maybe served as an alternative tool for clinical analysis.
College of Chemistry and Chemical Engineering, Central South University, Changsha, PR China.
Yong
Y
Huang
Ming
M
Shi
Shulin
S
Zhao
Hong
H
Liang
D000328:Adult; D046209:Electrophoresis, Microchip / Q000295:instrumentation / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D057230:Limit of Detection; D008163:Luminescent Measurements / Q000295:instrumentation / Q000379:methods*; D008297:Male; D008875:Middle Aged; D015203:Reproducibility of Results; D013739:Testosterone / Q000097:blood*
D013739:Testosterone