A novel colorimetric competitive aptamer assay for lysozyme detection based on superparamagnetic nanobeads.
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.
Université de Perpignan via Domitia, Laboratoire BAE, Building S 52 Av. Paul Alduy, 66860 Perpignan Cedex, France; Department of Biosciences and Biotechnology, Banasthali University, Rajasthan, 304022 India.
Rupesh K
RK
Mishra
Université de Perpignan via Domitia, Laboratoire BAE, Building S 52 Av. Paul Alduy, 66860 Perpignan Cedex, France; Interdisciplinary Research Centre in Biomedical Materials (IRCBM), COMSATS Institute of Information Technology (CIIT), Lahore 54000, Pakistan.
Akhtar
A
Hayat
Sabanci University Nanotechnology Research and Application Center, Orta Mahalle 34956, Tuzla, Istanbul, Turkey.
Geetesh K
GK
Mishra
Université de Perpignan via Domitia, Laboratoire BAE, Building S 52 Av. Paul Alduy, 66860 Perpignan Cedex, France.
Gaëlle
G
Catanante
Department of Biosciences and Biotechnology, Banasthali University, Rajasthan, 304022 India.
Vinay
V
Sharma
Université de Perpignan via Domitia, Laboratoire BAE, Building S 52 Av. Paul Alduy, 66860 Perpignan Cedex, France. Electronic address: jlmarty@univ-perp.fr.
Jean-Louis
JL
Marty
D052157:Aptamers, Nucleotide / Q000737:chemistry*; D015374:Biosensing Techniques / Q000379:methods*; D003124:Colorimetry / Q000379:methods*; D057230:Limit of Detection; D058185:Magnetite Nanoparticles / Q000737:chemistry*; D009113:Muramidase / Q000032:analysis*; D014920:Wine / Q000032:analysis*
D052157:Aptamers, Nucleotide; D058185:Magnetite Nanoparticles; D009113:Muramidase
Colorimetry; Competitive assay; Lysozyme; Superparamagnetic nanobeads; Wine