Kardiyak Miyozin Bağlayıcı Protein-C Tayini için Optik Temelli Sensörlerin Hazırlanması

Abstract

Cardiac myosin binding protein-C (cMyBP-C) plays a critical role in the contraction of cardiac muscle, and dysfunctions in this protein can lead to severe cardiovascular diseases. This thesis addresses the limitations of current diagnostic methods and the challenges encountered in diagnosing disorders related to cardiac myosin. It explores the potential of optical-based sensors in this field. The primary objective of the thesis is to develop and optimize surface plasmon resonance (SPR) sensors for the identification and analysis of cardiac myosin binding protein-C. In this context, molecularly imprinted polymer matrices are created using microcontact printing methods to recognize and bind specific molecular targets. The methods used to enhance the sensitivity and selectivity of the sensors through microcontact printing are discussed. These methods involve creating a polymeric mold that imprints the structure of the target molecule onto the sensor surface, thus facilitating the identification of the target molecule. This approach increases the likelihood of the target molecule binding to the sensor, thereby improving the sensor's performance. Microcontact printing methods enable the development of more reliable biological detection systems, which have broad applications in medical diagnostics, food safety, and environmental monitoring.

For the determination of cardiac myosin binding protein-C, kinetic analyses, selectivity studies, and various kinetic experiments in artificial plasma were conducted using cMyBP-C imprinted (MIP) and non-imprinted (NIP) poly(2-hydroxyethyl methacrylate-N-methacryloyl-L-tryptophan) (poly(HEMA-MATrp)) polymeric film-based SPR sensors. The SPR sensors prepared using the microcontact printing method for the cMyBP-C protein exhibited greater sensitivity compared to the non-imprinted SPR sensors. The sensitivity of both imprinted and non-imprinted sensors for detecting cMyBP-C was determined by preparing cMyBP-C solutions at different concentrations in pH 7.4 phosphate buffer and artificial plasma solutions, and adsorption kinetics were analyzed. The limit of detection (LOD) and limit of quantification (LOQ) for cMyBP-C imprinted SPR sensors were calculated to be 0.019 ng/mL and 0.064 ng/mL, respectively, based on the kinetic analysis data. To demonstrate the selectivity of the cMyBP-C imprinted sensors, competitive agents such as cardiac troponin T (cTnT) and creatine kinase-MB (CK-MB) were used. The results of the selectivity experiments showed that the cMyBP-C imprinted SPR sensors exhibited high selectivity and sensitivity. The findings of this study contribute to the development of new and effective sensors for the detection of cMyBP-C.