Ren Xiang, Yan Junrong, Wu Dan, Wei Qin, Wan Yakun
Key Laboratory of Interfacial Reaction & Sensing Analysis in Universities of Shandong, School of Chemistry and Chemical Engineering, University of Jinan , Jinan 250022, Shandong, China.
Institute of Life Sciences, Southeast University , Nanjing 210000, P.R. China.
ACS Sens. 2017 Sep 22;2(9):1267-1271. doi: 10.1021/acssensors.7b00495. Epub 2017 Sep 11.
Alzheimer's disease (AD) biomarkers can reflect the neurochemical indicators used to estimate the risk in clinical nephrology. Apolipoprotein E (ApoE) is an early biomarker for AD in clinical diagnosis. In this research, through bactrian camel immunization, lymphocyte isolation, RNA extraction, and library construction, ApoE-specific Nbs with high affinity were successfully separated from an immune phage display nanobody library. Herein, a colorimetric immunosensor was developed for the point-of-care testing of ApoE by layer-by-layer nanoassembly techniques and novel nanobodies (Nbs). Using highly oriented Nbs as the capture and detection antibodies, an on-site immunosensor was developed by detecting the mean gray value of fade color due to the glutaraldehyde@3-aminopropyltrimethoxysilane oxidation by HO. The detection limit of AopE is 0.42 pg/mL, and the clinical analysis achieves a good performance. The novel easily operated immunosensor may have potential application in the clinical diagnosis and real-time monitoring for AD.
阿尔茨海默病(AD)生物标志物可反映用于评估临床肾脏病风险的神经化学指标。载脂蛋白E(ApoE)是临床诊断中AD的早期生物标志物。在本研究中,通过双峰驼免疫、淋巴细胞分离、RNA提取和文库构建,从免疫噬菌体展示纳米抗体文库中成功分离出具有高亲和力的ApoE特异性纳米抗体(Nbs)。在此,通过逐层纳米组装技术和新型纳米抗体(Nbs)开发了一种用于ApoE即时检测的比色免疫传感器。使用高度定向的纳米抗体作为捕获和检测抗体,通过检测HO对戊二醛@3-氨丙基三甲氧基硅烷氧化导致的褪色平均灰度值,开发了一种现场免疫传感器。AopE的检测限为0.42 pg/mL,临床分析取得了良好的性能。这种新型的易于操作的免疫传感器可能在AD的临床诊断和实时监测中具有潜在应用。