In this study, it is aimed to determine cardiac troponin I by a surface plasmon resonance biosensor immobilized anti-cardiac troponin I monoclonal antibody. The immobilized anti-cardiac troponin I monoclonal antibody surface plasmon resonance biosensors were characterized with ellipsometry, atomic force microscopy and contact angle analysis. After that, surface plasmon resonance biosensor system was completed to biosensor system to investigate kinetic properties for cardiac tropinin I. The sensing ability of surface plasmon resonance biosensor was investigated with 0.001-8.0 ng/mL concentrations of cardiac tropinin I solutions. The limit of detection and limit of quantification were calculated as 0.00012 ng/mL and 0.00041 ng/mL, respectively. To show the selectivity of surface plasmon resonance biosensor competitive adsorption of cardiac tropinin I, myoglobin, immunoglobulin G and prostate specific antigen were investigated. Surface plasmon resonance biosensor was investigated five times with 0.5 ng/mL concentrations of cardiac tropinin I solution to show reuse of the chip. The results showed that surface plasmon resonance biosensor has high selectivity for cardiac tropinin I. The reproducibility of surface plasmon resonance sensors was investigated both on the same day and on different days for five times. To determine the usability, selectivity and validation studies of surface plasmon resonance biosensors were performed by enzyme-linked immunosorbent assay method.