期刊
ANALYTICAL METHODS
卷 6, 期 7, 页码 2233-2238出版社
ROYAL SOC CHEMISTRY
DOI: 10.1039/c3ay42075b
关键词
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资金
- Scientific Research Fund of Sichuan Provincial Education Department [11ZA231, 14ZA0303]
- Natural Science Foundation of Sichuan Province
- Key Project of Chengdu Normal University [CS13ZA02]
A portable and quantitative enzyme immunoassay with a glucometer readout was developed for the sensitive monitoring of neuron-specific enolase (NSE, as a model analyte) in a high-binding polystyrene 96-well microtiter plate (MTP), conjugated with monoclonal mouse anti-human NSE antibody (mAb(1)). Gold nanoparticles heavily functionalized with glucoamylase and polyclonal rabbit anti-human NSE antibody (pAb(2)) were utilized as the trace tag. A sandwich-type immunoassay format was adopted for the quantitative detection of NSE in the mAb(1)-functionalized MTP. Accompanying the gold nanoparticles, the carried glucoamylase could hydrolyze amylopectin in glucose. The produced glucose could be quantitatively monitored using a portable personal glucose meter (PGM). Under optimal conditions, the PGM-based immunoassay exhibited good analytical properties for the determination of the target NSE, and allowed the detection of NSE at concentrations as low as 0.05 ng mL(-1). The intra- and inter-assay coefficients of variation (CVs) were below 10% and 11%, respectively. The methodology was also evaluated by assaying 15 clinical serum samples, and showed good accordance between results obtained by the developed immunoassay and the referenced values.
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