The enzyme creatine kinase (CK) is one of the most well-established biomarkers in cardiovascular disease. For acute myocardial infarction (AMI) diagnosis, CK has a clinical comorbidity rate of 90%. This study has developed a novel electrochemical immunosensor using disposable graphite paper electrodes (GP). The GP electrodes were modified with gold nanoparticles (AuNP) to facilitate CK detection. Afterwards, GP electrodes were covalently immobilized with 6-mercapto-1-hexanol (6-MH), resulting in self-assembled monolayers (SAMs). Then, the electrodes were formed with a 3-Glycidyloxypropyltrimethoxysilane (3-GOPE) agent. Afterwards, the electrodes were immobilized with anti-CK (antibody creatine kinase) protein. In the final immobilization step, BSA (bovine serum albumin) protein was used to block non-covalent interactions.