抗hMOF单克隆抗体的制备与鉴定  

Preparation and Identification of Monoclonal Antibody Against hMOF

作　　者：孙厚良[1] 魏大鹏[2] 张小洪[2] 刘彦君[2] 章崇杰[2] 

机构地区：[1]重庆三峡医药高等专科学校免疫学教研室,万州404016 [2]四川大学华西基础医学与法医学院免疫学教研室,成都610041

出　　处：《四川大学学报（医学版）》2011年第5期721-724,共4页Journal of Sichuan University(Medical Sciences)

摘　　要：目的制备抗hMOF的单克隆抗体并对其生物学特性进行鉴定。方法以重组GST-hMOF融合蛋白作为免疫原免疫BALB/c小鼠,取其脾细胞与SP2/0细胞融合,经多次筛选及克隆化,建立可以稳定分泌抗hMOF单克隆抗体的杂交瘤细胞株。用ELISA、Western blot、免疫组化对此抗体特性进行鉴定。结果筛选到1株能稳定分泌抗hMOF单克隆抗体的细胞株4C1C8,亚类鉴定单抗为IgG1。ELISA法测定饱和硫酸铵盐析抗体效价为1∶409600,抗体相对亲和力为7.65×106L/mol;Western blot显示抗体能特异识别免疫原;细胞免疫组化显示此单抗可以特异的识别正常细胞株HL7702表达的hMOF。结论成功制备抗hMOF单克隆抗体,为进一步研究hMOF功能及机制奠定了基础。Objective To prepare and identify the monoclonal antibody（mAb） against hMOF.Methods BALB/C mice were immunized with protein from the spleen cells isolated and fused with SP2/0 cells.After several rounds of screening and cloning,the hybridoma cell strain secreting anti-hMOF mAb was obtained.Its specificity was evaluated with ELISA and Western blot,and the titer,immunoglobulin subtype and affinity of the mAb were measured. Results One cell strains of hybridoma were obtained and named as 4C1C8.The anti-hMOF mAb secreted by the hybridoma cell strain was identified as IgG1 subtype.The mAb titers in ascitic fluid were 1∶409600,as determined with ELISA with an affinity reaching to 7.65×106 L/mol.Western blot demonstrated that the antibodies could specifically recognize the immunogen.The cell immunohistochemistry proved that the antibody could recognize the hMOF antigens expressed on the normal cells HL7702.Conclusion The success in anti-hMOF mAb preparation provides the basis for further study of hMOF.

关 键 词：hMOF 单克隆抗体 制备与鉴定 

分 类 号：R392[医药卫生—免疫学]