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This paper aims to report on the development of a novel electrochemical amperometric immunosensor to diagnose early hepatocellular carcinoma (HCC) by detecting the Midkine (MDK) biomarker.

Anti-Midkine antibodies were immobilized covalently through carbodiimides chemistry on carbon screen-printed electrodes modified with carboxylated multi-walled carbon nanotubes. The development process was characterized using cyclic voltammetry, electrochemical impedimetric spectroscopy, Fourier transform infrared spectroscopy and atomic force microscopy. Differential pulse voltammetry was used to investigate the immunosensor performance in detecting MDK antigen within the concentration range of 1 pg/ml to 100 ng/ml.

MDK immunosensor exhibited high sensitivity and linearity with a detection limit of 0.8 pg/ml and a correlation coefficient of 0.99. The biosensor also demonstrated high selectivity, stability and reproducibility.

The developed MDK immunosensor could be a promising tool to diagnose HCC and reduce the number of related deaths.