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Abstract We present an advanced electrochemical immunosensor designed to detect the vascular endothelial growth factor (VEGF) precisely. The sensor is constructed on a modified porous gold electrode through a fabrication process involving the deposition of silver and gold on an FTO substrate. Employing thermal annealing and a de-alloying process, the silver is eliminated from the electrode, producing a reproducible porous gold substrate. Utilizing a well-defined protocol, we immobilize the heavy-chain (VHH) antibody against VEGF on the gold substrate, facilitating VEGF detection through various electrochemical methods. Remarkably, this immunosensor performs well, featuring an impressive detection limit of 0.05 pg/mL and an extensive linear range from 0.1 pg/mL to 0.1 µg/mL. This emphasizes it’s to measure biomarkers across a wide concentration spectrum precisely. The robust fabrication methodology in this research underscores its potential for widespread application, offering enhanced precision, reproducibility, and remarkable detection capabilities for the developed immunosensor.