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当前位置: 首页 > 文章 > 猪α干扰素基因的表达及其单克隆抗体的制备与鉴定 华中农业大学学报 2008,27 (3) 16-19
Position: Home > Articles > Prokaryotic Expression,Preparation and Characterization of Monoclonal Antibodies against Porcine IFN-α Journal of Huazhong Agricultural University 2008,27 (3) 16-19

猪α干扰素基因的表达及其单克隆抗体的制备与鉴定

作  者：
曾翠平;王晓杜;戴汉川;龙良启;伍晓雄
单  位：
华中农业大学动物医学院
关键词：
猪;α-干扰素;原核表达;单克隆抗体
摘  要：
将猪IFN-α(poIFN-α)成熟蛋白基因克隆于pET-28a载体中,转染大肠杆菌,并进行诱导表达。经SDS-PAGE检测,融合表达的蛋白poIFN-α大小约27 ku。收获融合表达的poIFN-α,免疫BALB/c小鼠,取脾细胞与SP2/0骨髓瘤细胞进行融合,通过间接ELISA方法对融合细胞进行检测,结果获得了1株能稳定分泌抗poIFN-α蛋白抗体的杂交瘤细胞株,命名为2H12株。Western-blotting结果及单克隆抗体亚型鉴定结果表明,单抗与重组蛋白发生特异性反应,单抗为IgG1亚型,且轻链为κ链。
译  名：
Prokaryotic Expression,Preparation and Characterization of Monoclonal Antibodies against Porcine IFN-α
作  者：
ZENG Cui-ping WANG Xiao-du DAI Han-chuan LONG Liang-qi WU Xiao-xiong (College of Veterinary Medicine,Huazhong Agricultural University,Wuhan 430070,China)
关键词：
porcine;IFN-α;prokaryotic expression;monoclonal antibody
摘  要：
The poIFN-α gene was cloned into a prokaryotic expression vector pET-28a,and a fusion expressed protein poIFN-α of 27 ku was obtained in E.coli.BALB/c mice were immunized intraperitoneally with purified poIFN-α protein.Murine myeloma cells SP2/0 were fused with the splenocytes of the immunized mice after the third immunization.An indirect ELISA was used to screen hybridomas for production of specific antibody in hybridoma cell.One hybridomas clone,named 2H12,which produced McAb steadily was obtained.The McAb showed strong reactivity with poIFN-α protein expressed by Western-blotting.The McAb belongs to IgG1 subtype,and its light chain was κ chain,which was subtyped by ELISA using acommercial kit.The McAb against by poIFN-α protein could be used for further analysis on the structure and function of poIFN-α.

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