Abstract
A homogeneous chemiluminescent immunoassay of thyroxine (T4) present in serum samples is described. The proposed method deployed the competitive immunoreaction of T4 and horseradish peroxidase (HRP)-labeled T4 (HRP–T4) with anti-T4 mouse monoclonal antibody (Ab). HRP–T4 and the HRP–T4–Ab complex were separated and quantified by using microchip electrophoresis (MCE) with chemiluminescence (CL) detection. The MCE separation was accomplished within 60 s. Highly sensitive CL detection was achieved by means of HPR-catalyzed luminol-H2O2 reaction. The linear range for T4 was 5–250 nM with a detection limit of 2.2 nM (S/N = 3).
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Bishop ML, Fody EP, Schoeff LE (eds) (2010) Clinical chemistry techniques, principles, correlations. Lippincott Williams & Wilkins, Philadelphia
Chin KP, Pin YC (2008) Heterophile antibody interference with thyroid assay. Intern Med 47:2033–2037
Ghosh S et al (2008) Interference in free thyroxine immunoassay. Eur J Intern Med 19:221–222
Gu J, Soldin OP, Soldin SJ (2007) Simultaneous quantification of free triiodothyronine and free thyroxine by isotope dilution tandem mass spectrometry. Clin Biochem 40:1386–1391
Chace DH et al (2009) Rapid metabolic and newborn screening of thyroxine (T4) from dried blood spots by MS/MS. Clin Chim Acta 403:178–183
Wu D, Qin J, Lin B (2008) Electrophoretic separations on microfluidic chips. J Chromatogr A 1184:542–559
Koutny LB et al (1996) Microchip electrophoretic immunoassay for serum cortisol. Anal Chem 68:18–22
Chiem N, Harrison DJ (1997) Microchip-based capillary electrophoresis for immunoassays: analysis of monoclonal antibodies and theophylline. Anal Chem 69:373–378
Bromberg A, Mathies RA (2004) Multichannel homogeneous immunoassay for detection of 2,4,6-trinitrotoluene (TNT) using a microfabricated capillary array electrophoresis chip. Electrophoresis 25:1895–1900
Wang J, Ibanez A, Chatrathi MP (2002) Microchip-based amperometric immunoassays using redox tracers. Electrophoresis 23:3744–3749
Wang J, Ibanez A, Chatrathi MP (2003) On-chip integration of enzyme and immunoassays: simultaneous measurements of insulin and glucose. J Am Chem Soc 125:8444–8445
Dishinger JF, Kennedy RT (2007) Serial immunoassays in parallel on a microfluidic chip for monitoring hormone secretion from living cells. Anal Chem 79:947–954
Phillips TM (2004) Rapid analysis of inflammatory cytokines in cerebrospinal fluid using chip-based immunoaffinity electrophoresis. Electrophoresis 25:1652–1659
Jin W, Li X, Gao N (2003) Simultaneous determination of tryptophan and glutathione in individual rat hepatocytes by capillary zone electrophoresis with electrochemical detection at a carbon fiber bundle—Au/Hg dual electrode. Anal Chem 75:3859–3864
Acknowledgments
Financial support from the National Natural Science Foundations of China (NSFC, Grant No. 20665002, 20875019 to SZ) and National Institutes of Health (SC1 GM089557) is gratefully acknowledged.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Zhao, S., Liu, YM. (2013). A Homogeneous Immunoassay of Thyroxine Based on Microchip Electrophoresis and Chemiluminescence Detection. In: Phillips, T., Kalish, H. (eds) Clinical Applications of Capillary Electrophoresis. Methods in Molecular Biology, vol 919. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-029-8_8
Download citation
DOI: https://doi.org/10.1007/978-1-62703-029-8_8
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-028-1
Online ISBN: 978-1-62703-029-8
eBook Packages: Springer Protocols