Lizozim Tayini İçin Lizozim Baskılanmış Yüzey Plazmon Rezonans Çipler

Abstract

Lyzozyme, a natural antimicrobial defense mechanism in the body, acts by breaking down the cell walls of microorganisms and is found in various tissues. Due to this property, it is considered an important component that protects the body from a range of pathogenic microorganisms. The detection of lysozyme is crucial for the diagnosis of certain diseases such as Alzheimer's and Crohn's. This thesis involves the use of molecular imprinting techniques to prepare lysozyme-imprinted surface plasmon resonance (SPR) sensors for the determination of lysozyme in aqueous solutions and artificial samples. In these studies, gold nanoparticles (AuNPs) are used, and the signal amplification provided by AuNPs is remarkable. Lyzozyme-imprinted polymeric film (MIP), lysozyme-imprinted gold nanoparticle-based polymeric film (AuNP MIP), and non-imprinted gold nanoparticle-based polymeric film (AuNP-NIP) surface plasmon resonance (SPR) sensors have been thoroughly examined through analyses and comparisons. The lysozyme detection sensitivity of imprinted and non-imprinted biosensors is investigated using lysozyme solutions (pH: 7.4 phosphate buffer), artificial urine, artificial tears, and artificial plasma. Sensors prepared with lysozyme imprinting and gold nanoparticles show higher sensitivity to lysozyme compared to non-imprinted and gold nanoparticle prepared sensors. Lysozyme solutions at different concentrations are used to determine the adsorption kinetics. Using kinetic analysis data from AuNP-MIP SPR sensors, the limit of detection (LOD) is calculated as 0.008 µg/mL, and the limit of quantification (LOQ) is 0.026 µg/mL. For MIP sensors, these values are determined as LOD 0.095 µg/mL and LOQ 0.317 µg/mL, respectively. To demonstrate the selectivity of lysozyme-imprinted AuNP-MIP and MIP sensors, competitive agents such as myoglobin and hemoglobin are used. Selectivity experiments show that lysozyme exhibits high selectivity and sensitivity for the prepared AuNP-MIP and MIP sensors. The results of this study contribute to the development of new and effective sensors in the field of lysozyme detection.