Journal
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
Volume 936, Issue -, Pages 74-79Publisher
ELSEVIER
DOI: 10.1016/j.jchromb.2013.08.002
Keywords
Chemiluminescence detection; Competitive immunoassay; Human serum; Microchip electrophoresis; Progesterone
Funding
- National Natural Science Foundations of China [21065002, 21175030]
- Natural Science Foundations of Guangxi Province [2012GXNS-FAA053024]
- BAGUI Scholar Program
- project of Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources (Guangxi Normal University), Ministry of Education of China
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A sensitive and rapid homogeneous immunoassay method based on microchip electrophoresis-chemiluminescence detection (MCE-CL) using luminol-hydrogen peroxide as chemiluminescence system catalyzed by horseradish peroxidase (HRP) was developed for the determination of progesterone (P). The assay was based on the competitive immunoreactions between HRP-labeled P antigen (HRP-P) and P with a limited amount of anti-P mouse monoclonal antibody (Ab), and MCE separation of free HRP-P and HRP-P-Ab immunocomplex followed by CL detection. The effect of various factors such as conditions for the CL reaction, MCE and incubation time for the immunoreactions were examined and optimized. Under optimal assay conditions, the MCE separation was accomplished within 80 s. The linear range of detection for P was 8-800 nM with a detection limit of 3.8 nM (signal/noise ratio = 3). This present method has been applied to determine Pin human serum samples from normal and pregnant women. The result indicates that the proposed MCE-CL based homogeneous immunoassay method can serve as an alternative tool for clinical assay of P. (C) 2013 Published by Elsevier B.V.
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