Li Yanxia, Li Yujun, Huang Lihua, Bin Qiu, Lin Zhenyu, Yang Huanghao, Cai Zongwei, Chen Guonan
Ministry of Education Key Laboratory of Analysis and Detection Technology for Food Safety (Fuzhou University), Fujian Province Key Laboratory of Analysis and Detection for Food Safety, Department of Chemistry, Fuzhou University, Fuzhou, 350002, China.
J Mater Chem B. 2013 Mar 7;1(9):1256-1262. doi: 10.1039/c2tb00398h. Epub 2013 Jan 18.
A novel molecular imprinting sensor based on TiO sol-gel layers embedding Cu(OH) was developed for protein recognition. Protein-imprinted TiO layers were prepared by liquid phase deposition (LPD) using lysozyme (Lys) as a template protein. The synthetic molecularly imprinted nanoparticles (MIPs) can catalyze o-phenylenediamine (OPDA) oxidation to produce fluorescence and color. Cu(OH) played a catalytic role in this reaction. The fluorescence intensity was related to the amount of MIPs, but has nothing to do with the imprinted protein. Taking advantage of this phenomenon, the imprinted protein can be detected using ELISA plates to fixed MIPs by OPDA oxidation. The fluorescence intensity at 568 nm (ex: 350 nm) was found to increase with increasing concentration of Lys. It was found that the limit of detection was 0.001 mg mL with the fluorescent sensor and 0.04 mg mL with the colorimetric sensor under optimized conditions. The results show that the MIPs reached saturated adsorption at 0.2 mg mL. Furthermore, the MIPs showed high selectivity compared to other tested proteins. In contrast, the fluorescence change of the non-imprinted TiO@Cu(OH) was only small. The proposed method will be a useful platform for recognition of protein with high sensitivity and specificity.
开发了一种基于嵌入Cu(OH)的TiO溶胶 - 凝胶层的新型分子印迹传感器用于蛋白质识别。以溶菌酶（Lys）作为模板蛋白，通过液相沉积（LPD）制备蛋白质印迹TiO层。合成的分子印迹纳米粒子（MIPs）可催化邻苯二胺（OPDA）氧化产生荧光和颜色。Cu(OH)在该反应中起催化作用。荧光强度与MIPs的量有关，但与印迹蛋白无关。利用这一现象，可通过ELISA板固定MIPs，利用OPDA氧化检测印迹蛋白。发现568nm（激发波长：350nm）处的荧光强度随Lys浓度的增加而增加。发现在优化条件下，荧光传感器的检测限为0.001mg/mL，比色传感器的检测限为0.04mg/mL。结果表明，MIPs在0.2mg/mL时达到饱和吸附。此外，与其他测试蛋白质相比，MIPs表现出高选择性。相比之下，非印迹TiO@Cu(OH)的荧光变化很小。所提出的方法将是一个用于高灵敏度和特异性识别蛋白质的有用平台。
