Eriş Şeyma, Çimen Duygu, Denizli Adil
Department of Chemistry, Hacettepe University, Ankara 06800, Turkey.
Bioengineering Division, Hacettepe University, Ankara 06800, Turkey.
ACS Omega. 2025 Jun 26;10(26):28055-28064. doi: 10.1021/acsomega.5c01607. eCollection 2025 Jul 8.
In this study, lysozyme-imprinted (Lyz-AuNP-MIP) and nonimprinted (AuNP-NIP) surface plasmon resonance (SPR) sensors modified with gold nanoparticles (AuNPs) were prepared for signal amplification and real-time determination of lysozymes from both aqueous lysozymes and artificial plasma, artificial urine, and artificial tear solutions. Furthermore, lysozyme-imprinted (Lyz-MIP) SPR sensors without AuNPs were prepared to study the signal enhancing effect of the AuNPs. The surface morphology of all prepared SPR sensors was characterized by atomic force microscopy and contact angle measurements. After the characterization study, kinetic studies were carried out using lysozyme solutions (pH 7.4 phosphate buffer), artificial plasma, artificial urine, and artificial tear prepared from 0.01-500 μg/mL concentrations using Lyz-AuNP-MIP SPR sensors for lysozyme determination. Using the kinetic analysis data obtained from the Lyz-AuNP-MIP SPR sensors, the limit of detection and limit of quantification were 0.008 and 0.026 μg/mL, respectively. Competitive agents, such as myoglobin and hemoglobin, were used to demonstrate the selectivity of Lyz-AuNP-MIP and AuNP-NIP SPR sensors. Furthermore, after optimizing the experimental studies for lysozyme determination, lysozyme determination was also performed in artificial plasma, artificial urine, and artificial tear solutions, and the recoveries were calculated to be approximately 99%.
在本研究中,制备了用金纳米颗粒(AuNPs)修饰的溶菌酶印迹(Lyz-AuNP-MIP)和非印迹(AuNP-NIP)表面等离子体共振(SPR)传感器,用于信号放大以及从水性溶菌酶和人工血浆、人工尿液及人工泪液溶液中实时测定溶菌酶。此外,还制备了不含AuNPs的溶菌酶印迹(Lyz-MIP)SPR传感器,以研究AuNPs的信号增强效果。通过原子力显微镜和接触角测量对所有制备的SPR传感器的表面形态进行了表征。在表征研究之后,使用Lyz-AuNP-MIP SPR传感器对浓度为0.01 - 500μg/mL的溶菌酶溶液(pH 7.4磷酸盐缓冲液)、人工血浆、人工尿液和人工泪液进行动力学研究以测定溶菌酶。利用从Lyz-AuNP-MIP SPR传感器获得的动力学分析数据,检测限和定量限分别为0.008和0.026μg/mL。使用肌红蛋白和血红蛋白等竞争剂来证明Lyz-AuNP-MIP和AuNP-NIP SPR传感器的选择性。此外,在优化了溶菌酶测定的实验研究后,还在人工血浆、人工尿液和人工泪液溶液中进行了溶菌酶测定,回收率计算约为99%。