Copper-Free Click Chemistry Assisted Antibodies Immobilization for Immunosensing Of IL-10 Cytokine

Copper-free variant of click chemistry has been successfully implemented to develop a new electrochemical immunosensor for cytokine interleukin-10 (IL-10) detection in saliva for heart failure (HF) monitoring. The approach involved the binding of dibenzocyclooctyne-amine (DBCO-NH2) onto the azide-thiol modified gold microelectrodes. The use of dibenzocyclooctynes decreased the activation energy for the cycloaddition click reaction, enabling its realization without the need for copper catalysis and with an efficiency greater than that of the copper catalysed ligation. Then, the biofunctionalization was performed through incubating the prepared sensor in the capture antibody of IL-10 (Anti-IL-10). The conjugation was formed via carbodiimide cross-linking and cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) techniques were used for characterizing step-by-step the modification of the gold microelectrode.