Ultrasensitive early detection of insulin antibody employing novel electrochemical nano-biosensor based on controllable electro-fabrication process

An ultrasensitive novel electrochemical nano-biosensor for rapid detection of insulin antibodies against diabetes antigens was developed in this research. The presence of insulin antibodies has been demonstrated to be a strong predictor for the development of type 1 diabetes in individuals who do not have diabetes but are genetically predisposed. The proposed nano-biosensor fabrication process was based on the optimized sequential electropolymerization of polyaniline and electrodeposition of gold nanoparticles on the surface of the functionalized gold electrode. The morphological and chemical characterization of the modified electrode was studied by field emission scanning electron microscopy (FE-SEM), energy dispersive X-ray spectroscopy (EDX), atomic force microscopy (AFM), Fourier transform infrared spectroscopy (FTIR), and micro Raman spectroscopy. Moreover, the role of each component in the modification of the electrode was studied by electrochemical methods systematically. After immobilizing insulin antigen and blocking with bovine serum albumin, the nano-biosensor was used for determining different concentrations of insulin antibody under the optimal conditions. This nano-biosensor could respond to insulin antibody with a linear calibration range from 0.001 ng ml-1 to 1000 ng ml-1 with the detection limit of 0.017 pg ml-1 and 0.034 pg ml-1 and selectivity of 18.544 μA ng-1 ml.cm-2 and 31.808 μA ng-1 ml.cm-2 via differential pulse voltammetry and square wave voltammetry, respectively. This novel nano-biosensor exhibited a short response time, high sensitivity, and good reproducibility. It was successfully used in determining the insulin antibody in human samples with a standard error of less than 0.178. Therefore, the nano-biosensor has the potential for the application of early detection of type 1 diabetes. To our best knowledge, label-free electrochemical detection of insulin antibody based on immunosensor is developed for the first time.