Çimen Duygu, Aslıyüce Sevgi, Tanalp Tuğçe Deniz, Denizli Adil
Hacettepe University, Department of Chemistry, Ankara, Turkey.
Gazi University, Vocational School of Health Services, Ankara, Turkey.
Anal Biochem. 2021 Nov 1;632:114221. doi: 10.1016/j.ab.2021.114221. Epub 2021 May 5.
In this study, a simple, fast, sensitive and selective surface plasmon resonance (SPR) sensor was prepared using molecular imprinting method for endotoxin detection. Endotoxin imprinted and non-imprinted poly(hydroxyethyl methacrylate-N-methacryloyl-(L)-histidine methyl ester) based nanofilms were synthesized on the SPR chip surfaces using ultraviolet polymerization. Endotoxin imprinted and non-imprinted SPR sensors were characterized by using contact angle, atomic force microscopy and ellipsometry. After characterization studies, kinetic studies was carried out in the concentration range of 0.5-100 ng/mL. The limit of detection and quantification were obtained as 0.023 and 0.078 ng/mL, respectively. The response time for the equilibration, adsorption and regeneration was approximately 14 min. The selectivity studies with cholesterol and hemoglobin of endotoxin imprinted SPR sensor were examined. Validation studies were carried out via limulus amebocyte lysate (LAL) in order to demonstrate the applicability of the SPR sensor.
在本研究中,采用分子印迹法制备了一种用于内毒素检测的简单、快速、灵敏且具有选择性的表面等离子体共振(SPR)传感器。通过紫外光聚合在SPR芯片表面合成了基于聚(甲基丙烯酸羟乙酯-N-甲基丙烯酰基-(L)-组氨酸甲酯)的内毒素印迹和非印迹纳米薄膜。利用接触角、原子力显微镜和椭偏仪对印迹和非印迹的SPR传感器进行了表征。表征研究之后,在0.5-100 ng/mL的浓度范围内进行了动力学研究。检测限和定量限分别为0.023和0.078 ng/mL。平衡、吸附和再生的响应时间约为14分钟。对内毒素印迹SPR传感器与胆固醇和血红蛋白的选择性进行了研究。为了证明SPR传感器的适用性,通过鲎试剂(LAL)进行了验证研究。