The enzyme creatine kinase (CK) is a biomarker that plays an extremely significant role in the early detection of cardiovascular disorders. Serum levels of CK are regularly monitored in patients with heart attacks, one of the most critical cardiovascular illnesses. In this study, a highly sensitive electrochemical immunosensor system was designed for the importance of early diagnosis of CK. This immunosensor system was developed by immobilizing 11- mercaptoundecanoic acid (11-MuA) on disposable indium tin oxide-polyethylene terephthalate (ITO-PET) electrodes. Electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV) and single frequency impedance (SFI) techniques were utilized throughout the immobilization process during the construction of the immunosensor. In addition, the proposed CK immunosensor system involves thorough analytical research, which may include linear determination range, repeatability, reproducibility, square wave voltammetry, storage capability, and regeneration. The suggested immunosensor was also characterized using scanning electron microscopy (SEM). The proposed immunosensor system demonstrated a broad dynamic range (0.1 pg/mL - 100 pg/mL), as well as a low limit of detection (LOD) and a low limit of quantification (LOQ) of 0.018 pg/mL and 0.0394 pg/mL, respectively. Finally, the immunosensor was tested on human serum samples, proving that it could be utilized in clinical situations.