期刊
BIOSENSORS & BIOELECTRONICS
卷 26, 期 2, 页码 517-522出版社
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2010.07.060
关键词
Testosterone; Immunosensor; Electrochemical; Magnetic particles; Screen-printed electrodes
类别
资金
- Ministerio de Ciencia e Innovacion [DPS2008-07005-C02-01, CTQ2009-12650]
- Comunidad de Madrid [S2009/PPQ-1642]
A disposable electrochemical immunosensor using screen-printed carbon electrodes (SPCEs) and protein A-functionalized magnetic beads (MBs) was developed for the determination of testosterone. Anti-testosterone was immobilized onto MBs and a direct competitive immunoassay involving testosterone labeled with peroxidase (HRP) was performed. The resulting conjugate was trapped on the SPCE with a small magnet. Testosterone determination was carried out by amperometry at -0.2 V upon H2O2 additions using hydroquinone (HQ) as the redox mediator. The experimental variables involved in the immunosensor response to testosterone were evaluated. Under the optimized conditions, a calibration plot for testosterone was obtained with a linear range between 5.0 x 10(-3) and 50 ng/mL (r = 0.995). The detection limit was 1.7 pg/mL and the EC50 was 0.25 +/- 0.04 ng/mL. These characteristics are notably better than those achieved with other reported immunosensors. Furthermore, anti-testosterone/MBs conjugates were shown to be stable for at least 25 days. A good selectivity was also found against other steroid hormones. The usefulness of the immunosensor was demonstrated by analyzing human serum spiked with 1 and 10 ng/mL testosterone. (C) 2010 Elsevier B.V. All rights reserved.
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