OVA-CdTe-ZnSe核壳量子点探针制备及其应用.docVIP

OVA-CdTe/ZnSe核壳量子点探针制备及其应用[摘要] 目的：制备CdTe/ZnSe核壳量子点标记卵清白蛋白（OVA）探针OVA-CdTe/ZnSe，并检测其特异性荧光示踪作用。方法：将CdTe/ZnSe核壳量子点与OVA偶联制备荧光探针，通过凝胶电泳和光谱分析鉴定偶联效果；然后示踪OVA致敏的小鼠外周血嗜碱粒细胞（BASO）表面IgE高亲和力受体FcεRI。结果：偶联OVA后的量子点分子量增加，荧光增强且荧光峰位从628 nm红移至635 nm。通过激光共聚焦显微镜能清晰观测到OVA-CdTe/ZnSe探针对BASO表面IgE-FcεRI复合物的荧光标记。结论：OVA-CdTe/ZnSe探针能示踪OVA特异性IgE-FcεRI复合物，此探针在OVA致敏动物模型的特异性IgE检测中具有一定应用价值。 [关键词] CdTe/ZnSe核壳量子点；免疫荧光标记；鸡卵清白蛋白；IgE [中图分类号] R392-3 [文献标识码] A [文章编号] 1673-7210（2011）11（c）-019-03 Preparation and fluorescence-imaging application of OVA-CdTe/ZnSe core/shell quantum dots NI Bin, HU Huajun* Department of Pharmacy, College of Life Science, China Jiliang University, Zhejiang Province, Hangzhou 310018, China [Abstract] Objective: To establish a method to detect the IgE-FcεRI on BASO cells by using CdTe/ZnSe core/shell quantum dots(QDs) immuno-fluorescence labeling technology. Methods: CdTe/ZnSe core/shell QDs were coupled to the OVA with N-hydroxysuccinimide (NHS). After confirmation by agarosegel electrophoresis method and spectra analysis, the coupled product (OVA-CdTe/Znse) was used to detect the IgE-FcεRI on BASO cells. Results: Spectra analysis showed that the fluorescence intensity of OVA-CdTe/Znse was enhanced and the emission peak underwent a red-shift from 628 nm to 635 nm. Confocal fluorescence microscopy further showed that OVA-IgE-FcεRI on BASO cells characterized by the red fluorescence. Conclusion: OVA-CdTe/Znse fluorescence probes can effectively recognize IgE-FcεRI on BASO cells, which can be used in allergic animal model for detecting OVA specific IgE. [Key words] CdTe/ZnSe core/shell quantum dots; Immuno-fluorescence labeling; OVA; IgE 支气管哮喘（bronchial asthma，简称哮喘）属于过敏性疾病的一种，其发病机制复杂，目前全球约有3亿哮喘患者，被WHO列为四大顽症之一。鉴于人体试验的局限性，对哮喘的干预研究往往通过建立与人相似症状的动物模型来进行。常用的哮喘动物（大鼠、小鼠、豚鼠）模型都存在不足之处。如豚鼠致敏后的变态反应与人类的不同，其更多由IgG介导；大鼠、小鼠制作的哮喘模型能产生特异性IgE，可临床指征不明显[1]。因此有必要寻找和开发新的哮喘动物模型。但哮喘新模型的开发还面临一个具体问题，即除大鼠、小鼠、豚鼠外的其他实验动物在卵清白蛋白（OVA）致敏后，因缺乏血清OVA特异性IgE的检测试剂，使致敏效果难以判断，这也阻碍了新模型的开发[2]。根据外周血嗜碱粒细胞（