Abstract
An aptamer-based method is described for electrochemical determination of lysozyme. A glassy carbon electrode was modified with a nanocomposite composed of multi-walled carbon nanotubes, poly(diallyl dimethyl ammonium chloride), and carbon quantum dots. The composition of the nanocomposite (MWCNT/PDDA/CQD) warrants good electrical conductivity and a high surface-to-volume ratio. The lysozyme-binding aptamers were immobilized on the nanocomposite via covalent coupling between the amino groups of the aptamer and the carboxy groups of the nanocomposite. The modified electrode was characterized by electrochemical impedance spectroscopy, cyclic voltammetry and differential pulse voltammetry. The use of this nanocomposite results in a considerable enhancement of the electrochemical signal and contributes to improving sensitivity. Hexacyanoferrate was used as an electrochemical probe to study the dependence of the peak current on lysozyme concentration. In the presence of lysozyme, the interaction of lysozyme with immobilized aptamer results in a decrease of the peak current, best measured at +0.15 V vs. Ag/AgCl. A plot of peak current changes versus the logarithm of the lysozyme concentration is linear in the 50 fmol L-1 to 10nmol L-1 concentration range, with a 12.9 fmol L-1 detection limit (at an S/N ratio of 3). The method is highly reproducible, specific and sensitive, and the electrode has a rapid response. It was applied to the determination of lysozyme in egg white, serum, and urine. Graphical abstract Schematic of a nanocomposite composed of multi-walled carbon nanotubes (MWCNTs), poly(diallyldimethyl ammonium chloride) (PDDA), and carbon quantum dots (CQDs) for use in a lysozyme aptasensor. The aptamer wasimmobilized on the surface, and bovine serum albumin (BSA) was applied to block the surface. The changes of peak current for the electrochemical probehexacyanoferrate (Fe(CN)63-/4-) in the presence and absence of lysozyme was traced.
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