In this work, a novel, sensitive, and rapid electrochemical biosensor was employed to detect lysozyme (Lys) using a double receptor of molecular imprinted polymer (MIP)-aptamer. First, a glassy carbon electrode (GCE) was modified with a nanocomposite consisting of multi-wall carbon nanotubes (MWCNTs), nitrogen-doped carbon quantum dots (N-CQDs), and chitosan. Subsequently, aptamer (Apt)-Lys complex was immobilized on MWCNTs-N-CQDs-chitosan/GCE via binding between carboxyl groups present in the nanocomposite and the terminal amine groups of the aptamer. Following that, methylene blue monomer was electrochemically polymerized around the Apt-Lys complex on the MWCNTs-N-CQDs-chitosan/GCE surface. Finally, after the template removal, the remaining cavities along with the aptamers created a new hybrid receptor of MIP-aptamer. The MWCNTs-N-CQDs-chitosan nanocomposite could provide large amounts of carboxyl groups for binding to amino-functionalized aptamers, considerable electrical conductivity, and a high surface-to-volume ratio. These beneficial features facilitated the Apt-Lys complex immobilization and gave improved electrochemical signal. The obtained MIP-aptamer hybrid receptor allowed lysozyme determination even at concentrations as low as 4.26 fM within the functional range of 1 fM to 100 nM.
Keywords: Electrochemical biosensor; Lysozyme; MIP-aptamer hybrid receptor; Methylene blue; Nitrogen-doped carbon quantum dots.
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