抗淀粉样蛋白寡聚体单链抗体融合蛋白在真核细胞中表达的初.PDFVIP

安徽农业大学学报, 2013, 40(4): 685-689 Journal of Anhui Agricultural University 网络出版时间：2013-7-10 18:25:34 [URL] /kcms/detail/34.1162.S1825.006.html 抗淀粉样蛋白寡聚体单链抗体融合蛋白 在真核细胞中表达的初步研究 1 1* 2 2* 周维维 ，汪维云 ，刘瑞田 ，于晓琳 (1. 安徽农业大学生命科学学院，合肥 230036 ；2. 中国科学院过程工程研究所，生化工程国家重点实验室，北京 100190) 摘 要：阿尔茨海默病（Alzheimer’s disease, AD ）是一种由脑内淀粉样蛋白（Aβ）聚集引起的，以老年人记 忆和认知功能丧失为临床表征的神经退行性疾病。研究表明，Aβ寡聚体是 AD 发生发展的主要因素。应用自行筛 选出的能够特异识别淀粉样蛋白寡聚体的单链抗体 W20 基因，构建了以绿色荧光蛋白为报告基因的真核表达载体 pSNAV2.0-W20-EGFP ，并应用流式细胞分选、蛋白免疫印迹以及荧光显微镜等技术对该单链抗体融合蛋白在真核 细胞中的表达进行了初步研究，为针对AD 的抗体基因治疗奠定了基础。 关键词：淀粉样蛋白；单链抗体（scFv ）；阿尔茨海默病 中图分类号：Q51 文献标识码：A 文章编号：1672−352X (2013)04−0685−05 Expression of single-chain variable fragment antibody specifically recognizing beta-amyloid oligomers in 293T cells 1 1 2 2 ZHOU Wei-wei , WANG Wei-yun , LIU Rui-tian , YU Xiao-lin (1. School of Life Science, Anhui Agricultural University, Hefei 230036 ； 2. National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, China Academy of Sciences, Beijing 100 190) Abstract: Alzheimer’s disease is the most common form of neurodegenerative disease that leads to a pro- gressive loss of memory and cognitive functions. Increasing evidence indicates that beta-amyloid oligomers are pivotal toxins in the pathogenesis of Alzheimers disease. In this study, W20, a single-chain variable fragment antibody specifically recognizing beta-amyloid oligomers was cloned to pSNAV2.0-EGFP vector and its ex- pression in 293T cells was examined by flow cytometry, western blot and fluorescence microscope. Such scFv antibody eukaryotic expression vectors may