A novel colorimetric competitive aptamer assay for lysozyme detection based on superparamagnetic nanobeads

被引:34
作者
Mishra, Rupesh K. [1 ,3 ]
Hayat, Akhtar [1 ,2 ]
Mishra, Geetesh K. [4 ]
Catanante, Gaelle [1 ]
Sharma, Vinay [3 ]
Marty, Jean-Louis [1 ]
机构
[1] Univ Perpignan, Lab BAE, Via Domitia,Bldg S 52 Av Paul Alduy, F-66860 Perpignan, France
[2] CIIT Ctr Hlth Res, IRCBM, Lahore 54000, Pakistan
[3] Banasthali Univ, Dept Biosci & Biotechnol, Vanasthali 304022, Rajasthan, India
[4] Sabanci Univ, Nanotechnol Res & Applicat Ctr, TR-34956 Istanbul, Turkey
关键词
Lysozyme; Wine; Superparamagnetic nanobeads; Colorimetry; Competitive assay; EGG-WHITE LYSOZYME; DNA; APTASENSOR; NANOPARTICLES; AMPLIFICATION; IMMUNOASSAY; ELECTRODES; PROTEINS; RESIDUES; SAMPLES;
D O I
10.1016/j.talanta.2016.12.083
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Lysozyme (Lys) commonly presents in wines and are known to cause toxicological impact on human health. The need of highly sensitive and reliable detection methods are evident in such matrix. In this work, we developed a competitive aptamer based assay for detection of Lys by employing carboxylated magnetic beads as a support to immobilize the target molecule Lys. The used aptamer sequence was biotinylated which further binds with Streptavidin-Alkaline phosphatase (Stp-ALP) in the micro wells. Colorimetric tests were performed in order to optimize different experimental parameters. The Lys assay showed a good linearity in the range of 5-140 nM with a limit of detection (LOD) 10 nM. The mid-point value (IC50) 110 nM and the analysis time (60 min) validated the developed aptasensor as a promising tool for routine use. The assay displayed good recoveries of Lys in the range 99.00-99.27% and was demonstrated for the detection of Lys in wine samples.
引用
收藏
页码:436 / 441
页数:6
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