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.
出处
《免疫学杂志》
CAS
CSCD
北大核心
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)
