KGF-2单克隆抗体间接ELISA筛选方法的建立  

Establishment of Indirect ELISA for Preparing the Monoclonal Antibody against KGF-2

作　　者：朱佳男[1,2] 姚娜[2] 官丽莉[2] 车莹[1,2] 李校堃[2] 

机构地区：[1]吉林农业大学研究生学院,吉林长春130118 [2]吉林农业大学生物反应器与药物开发教育部工程研究中心,吉林长春130118

出　　处：《北华大学学报（自然科学版）》2010年第3期222-224,共3页Journal of Beihua University（Natural Science）

基　　金：科技部重大新药创制项目(2008ZX09101)

摘　　要：目的建立KGF-2单克隆抗体间接ELISA的筛选方法 ,制备KGF-2单克隆抗体.方法以KGF-2为抗原免疫BALB/c小鼠,获得抗血清.通过棋盘滴定方法确定间接ELISA的抗原包被浓度及一抗、二抗最佳工作浓度;利用建立特异性好的间接ELISA方法筛选杂交瘤细胞株.结果通过棋盘滴定最终确定了500μg/L的抗原包被浓度、1∶1 000的一抗血清稀释倍数及1∶2 000的二抗血清稀释倍数;筛选到两株可稳定分泌抗体、效价较高的杂交瘤细胞株.结论该研究建立了灵敏度高、特异性好的间接ELISA筛选体系,可用于KGF-2单克隆抗体的制备及检测.Objective To prepare the monoclonal antibody against KGF-2 and develop an indirect competitive ELISA.Method Present anti-serum from BALB/c mice by injecting KGF-2 antigen.Confirm the best consistence of the antigen,the first antibody and the second antibody with method of square titration.McAb hybridoma was screened with the method of indirect ELISA.Results Based on the result of the square titration,antigen presidium consistence was 500μg/L;first antibody and second antibody should be diluted to 11000 and 12000.Two hybridoma cells secreted McAb against KGF-2 was screened.Conclusion The method of indirect ELISA was set up effectively and reliably,which could be used to screen and present KGF-2 McAb.

关 键 词：KGF-2单克隆抗体 棋盘滴定 杂交瘤细胞 

分 类 号：R392.11[医药卫生—免疫学]