Cross-linked chitosan/thiolated graphene quantum dots as a biocompatible polysaccharide towards aptamer immobilization

Chitosan has a number of commercial and possible biomedical uses. Chitosan as a polysaccharide is a bioactive polymer with a variety of applications due to its functional properties such as antibacterial activity, non-toxicity, ease of modification, and biodegradability. In this work, cross-linked chitosan/thiolated graphene quantum dot as a biocompatible polysaccharide was modified by gold nanoparticle and used for immobilization of ractopamine (RAC) aptamer. A highly specific DNA-aptamer (5'-SH-AAAAAGTGCGGGC-3'), selected to RAC was immobilized onto thiolated graphene quantum dots (GQDs)-chitosan (CS) nanocomposite modified by gold nanostructures (Au NSs) and used for quantification of RAC. Different shapes of gold nanostructures with various sizes from zero-dimensional nanoparticles to spherical structures were prepared by one-step template-assistant green electrodeposition method. Fully electrochemical methodology was used to prepare a new transducer on a glassy carbon surface which provided a high surface area to immobilize a high amount of the aptamer. Therefore, a label free electrochemical (EC) apta-assay for ultrasensitive detection of RAC was developed. A special immobilization media consisting of Au NSs/GQDs-CS/Cysteamine (CysA) was utilized to improve conductivity and performance of the biosensor. The RAC aptamer was attached on the Au NSs of the composite membrane via AuS bond. The fabrication process of the EC aptamer based assay was characterized by some electrochemical techniques. The peak currents obtained by differential pulse voltammetry decreased linearly with the increasing of RAC concentrations and the apta-assay responds approximately over a wide dynamic range of RAC concentration from 0.0044 fM to 19.55 μM. The low limit of quantification was 0.0044 fM.