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

• Published in: Talanta (Oxford) 2017-04, Vol.165, p.436-441
• Main Authors: Mishra, Rupesh K., Hayat, Akhtar, Mishra, Geetesh K., Catanante, Gaëlle, Sharma, Vinay, Marty, Jean-Louis
• Format: Article
• Language: English
• Subjects: Aptamers, Nucleotide - chemistry; Biosensing Techniques - methods; Chemical Sciences; Colorimetry; Colorimetry - methods; Competitive assay; Environmental Sciences; Life Sciences; Limit of Detection; Lysozyme; Magnetite Nanoparticles - chemistry; Muramidase - analysis; Superparamagnetic nanobeads; Wine; Wine - analysis
• ISSN: 0039-9140; 1873-3573
• DOI: 10.1016/j.talanta.2016.12.083

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 5–140nM with a limit of detection (LOD) 10nM. The mid-point value (IC50) 110nM and the analysis time (60min) 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. [Display omitted] •Highly sensitive lysozyme detection route.•First report on MB coupled aptamer assay.•Assay based on unique combination of biotinylated aptamer and Streptavidin interaction.