A direct and simple immobilization route for immunosensors by CNBr activation for covalent attachment of anti-leptin: obesity diagnosis point of view

Leptin is a peptide hormone produced in adipose tissue that works as an antiobesity hormone by balancing energy intake and expenditure. We aimed to develop an ultrasensitive electrochemical immunosensor based on a novel immobilization technique for the early detection of leptin-related diseases in this work. Although several methods for immobilizing antibodies to the biosensor recognition element are known, it is necessary to utilize novel, cost-effective, and less complicated immobilization procedures. When compared with currently utilized immobilization techniques for leptin measurement, this novel method is more efficient, easy to prepare, and sensitive, with a broad detection range. Indium tin oxide-coated polyethylene terephthalate (ITO-PET) sheets were used as the working electrode. ITO-PET sheets were modified using cyanogen bromide (CNBr) to immobilize the anti-leptin antibody through covalent interactions. Each stage of the proposed biosensors was characterized by electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) methods, and extensive characterization studies were carried out. The designed biosensor has a wide linear detection range (0.05–100 pg/mL), low limits of detection (LOD) (0.0086 pg/mL) and quantification (LOQ) (0.0287 pg/mL). It was concluded that although it is disposable, the ITO-PET working electrode retains its activity even in repeated studies. In addition, the new immobilization procedure provided by CNBr for the designed biosensor fabrication can be effectively used in other biosensing applications.