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糖基化纳米粒子的合成及其在唾液酸结合蛋白检测中的应用
祁晓晓1, 李学兵2, 周绪正1, 张继瑜1, 张振兴2, 吴培星1
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(1.中国农业科学院 兰州畜牧与兽药研究所, 兰州 730050;2.中国科学院 微生物研究所/病原微生物与免疫学重点实验室, 北京 100101)
摘要:
为实现流感病毒的简便快速检测,建立一种基于糖基化纳米粒子探针的荧光共振能量转移(FRET)检测方法。采用配体交换法,将合成的唾液酸寡糖链修饰到量子点和金纳米粒子表面,制备出了糖基化量子点和糖基化金纳米粒子,并对其理化性质进行了表征。结果表明:这2种纳米材料均具有良好的水溶性和光学特性,可分别作为能量供体和受体探针用于FRET检测体系;建立了一种基于FRET的唾液酸结合蛋白检测体系,麦胚凝集素(WGA),最低可检测浓度为4 nmol/L,证明该体系可快速(5 min)且灵敏地检测这种唾液酸结合蛋白。由于流感病毒表面的血凝素(HA)可特异性识别并结合唾液酸寡糖,本研究为进一步研发基于FRET的简单、快速的流感病毒检测方法奠定了基础。
关键词:  唾液酸结合蛋白  荧光共振能量转移  量子点  糖基化纳米粒子  麦胚凝集素
DOI:10.11841/j.issn.1007-4333.2019.12.10
投稿时间:2019-04-10
基金项目:国家现代农业产业技术体系(CARS-37);国家自然科学基金面上项目(31770862)
Construction of glycan-functionalized nanoparticles and its application in the detection of sialic acid-binding protein
QI Xiaoxiao1, LI Xuebing2, ZHOU Xuzheng1, ZHANG Jiyu1, ZHANG Zhenxing2, WU Peixing1
(1.Lanzhou Institute of Husbandry and Pharmaceutical, Chinese Academy of Agricultural Sciences, Lanzhou 730050, China;2.Institute of Microbiology/Key Laboratory of Pathogenic Microbiology and Immunology, Chinese Academy of Sciences, Beijing 100101, China)
Abstract:
For the rapid and simple detection of influenza viruses,a novel strategy based on fluorescence resonance energy transfer (FRET) was developed.By using ligand exchange method,glycan-functionalized quantum dots (gQDs) and glycan-functionalized gold nanoparticles (gAuNPs) were synthesized,respectively.The results showed that:The characterization of gQDs and gAuNPs revealed that they could be used as an effective FRET donor-acceptor pair because of their excellent optical properties,high aqueous solubility and stability;Based on these two glycan-functionalized nanomaterials,a FRET biosensor system was developed.Wheat germ agglutinin,which is a sialic acid-binding protein (SABP),was detected within 5 minutes with detection limit of 4 nmol/L.Because influenza viruses use their envelope SABP,hemagglutinin,to specificly recognize and attach to sailic acids displayed on the host cell surface.The method developed in this study would provide bases to develop gQDs and gAuNPs-based FRET systems for rapid and simple influenza virus detection.
Key words:  sialic acid-binding protein  fluorescence resonance energy transfer  quantum dots  gold nanoparticles  wheat germ agglutinin