平榛主要过敏原Corh1单克隆抗体的制备与鉴定  

Preparation and identification of monoclonal antibodies against Cor h1 allergen

作　　者：赵郭存[1] 张强[1] 邹菊[1] 黄钟[1] 刘志刚[1] 

机构地区：[1]深圳大学过敏反应与免疫学研究所,518060

出　　处：《免疫学杂志》2011年第6期523-526,共4页Immunological Journal

基　　金：国家自然科学基金(30871752);深圳市深港创新圈项目(CXQ2008026);深圳大学创新科研团队基金(200904)

摘　　要：目的制备和鉴定鼠抗平榛主要变应原Cor h 1的单克隆抗体(Monoclonal Antibody,McAb)。方法用重组Cor h 1蛋白为免疫原,免疫Balb/c小鼠,取免疫小鼠脾细胞与小鼠骨髓瘤NS-1细胞融合。通过间接ELISA法筛选分泌特异性McAb的杂交瘤细胞。用杂交瘤细胞株诱导小鼠产生腹水,再用蛋白A亲和层析法纯化抗体。采用Ig类与亚类鉴定试剂盒鉴定该单克隆抗体的Ig亚型;通过间接ELISA、Western Blotting鉴定该McAbs的特性和交叉性。结果获得4株可稳定分泌鼠抗平榛主要变应原Cor h 1的McAbs,其Ig亚型均为IgG1,均具有良好的效价;ELISA和Western Blotting分析表明该4株单抗均能识别重组Cor h 1蛋白,其中3株单抗能够识别天然平榛提取物。结论成功制备了4株鼠抗平榛主要变应原Cor h 1的单克隆抗体,为建立平榛主要变应原Cor h 1检测及纯化方法奠定了基础。This stuffy ained to prepare monoclonal antibodies against Cor h 1 allergen from Dermatophagoides farinae and to characterize their properties.Balb/c mice were immunized with purified recombinant Cor h 1 allergen protein,and the splenocytes of the immunized mice were fused with NS-1 myeloma cells by hybridoma technique.The McAbs against Cor h 1 allergen were purified using affinity chromatography on immobilized Protein A,and identified by ELISA and Western blotting for the specificity,subtype,titers and cross-reactivity.Four hybridoma cell lines secreting McAbs against Cor h 1 allergen were obtained,which were determined as IgG1 subtype and possessed good ascites titers.The four McAbs against Cor h 1 allergen all recognized recombinant Cor h 1 allergen,and the three of them also recognized native extract from Corylus heterophylla.In our study,four monoclonal antibodies against Cor h 1 allergen have prepared successfully,which will facilitate the establishment of detection method for Cor h 1 allergen.

关 键 词：Corh1 主要变应原 单克隆抗体 杂交瘤细胞 交叉性 

分 类 号：R392.11[医药卫生—免疫学]