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CE immunoassay with enhanced chemiluminescence detection of erythropoietin using silica dioxide nanoparticles as pseudostationary phase
Author(s) -
Wang Wenjun,
Zhang Sichun,
Liu Chenghui,
Lu Lingzhao,
Wang Shidong,
Zhang Xinrong
Publication year - 2009
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200800616
Subject(s) - detection limit , chemiluminescence , immunoassay , chromatography , chemistry , analyte , phosphate buffered saline , buffer (optical fiber) , nanoparticle , buffer solution , silicon dioxide , materials science , nanotechnology , metallurgy , antibody , immunology , telecommunications , computer science , biology
The measurement of serum erythropoietin (EPO) is important for the detection of recombinant human EPO misuse in sports. In this paper, we have developed a sensitive and rapid CE immunoassay with enhanced chemiluminescence detection of EPO, in which silica dioxide nanoparticles (SiO 2 NPs) were used as the pseudostationary phase to improve the separation efficiency of analytes. By adopting SiO 2 NPs in the CE immunoassay, the separation can be successfully performed in neutral running buffer solution. However, running buffer of extreme pH was still necessary for the conventional CZE mode. Neutral phosphate buffer (pH 7.4) containing SiO 2 NPs and poly(ethylene oxide) (PEO) was chosen as running buffer. The influences of SiO 2 NPs, PEO and buffer concentration on the separation efficiency and EPO detection were investigated. EPO‐horse radish peroxidase (HRP) and immunocomplex were baseline separated in a 10 mM phosphate buffer (pH 7.4) consisting of 0.08% PEO and 0.08% SiO 2 NPs. The linear range for EPO was 1.8–158.0 ng/mL and the detection limit was 0.9 ng/mL. The assay was successfully applied for the quantification of EPO in human sera and the results correlated well with those obtained using chemiluminescence immunoassay kits, thus demonstrating that the present method was a potential powerful tool for EPO misuse detection and clinical diagnosis.