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当前位置: 首页 > 文章 > 盐酸金霉素人工抗原的合成及鼠源多抗血清的制备 河南农业科学 2012,41 (7) 138-141
Position: Home > Articles > Synthesis of Chlortetracyline Artificial Antigen and Preparation of Its Mice Polyclonal Antiserum Journal of Henan Agricultural Sciences 2012,41 (7) 138-141

盐酸金霉素人工抗原的合成及鼠源多抗血清的制备

作  者：
李靓;职爱民;李青梅;刘庆堂;侯玉泽;胡骁飞;邓瑞广;张改平
单  位：
河南科技大学食品与生物工程学院;河南省农业科学院农业部动物免疫学重点开放实验室
关键词：
盐酸金霉素;多抗血清;人工抗原
摘  要：
采用碳二亚胺法将金霉素(chlortetracyline,CTC)偶联到载体蛋白BSA(牛血清白蛋白)和OVA(鸡卵清蛋白)上,形成免疫原BSA-CTC与包被原OVA-CTC。通过紫外分光光度法和聚丙烯酰胺凝胶电泳法进行抗原鉴定,用间接ELISA测定多抗血清(pAb)效价,阻断ELISA鉴定其敏感性。结果表明,在UV下出现了明显特征峰,BSA-CTC的峰与BSA、CTC相比均发生改变,表明半抗原与载体成功偶联。共免疫了5只小鼠,pAb效价均达到1×10-3,其中3号小鼠多抗血清抑制效价最低,IC50(半数抑制质量浓度)为71.47ng/mL。由此得出,获得了高效价、强特异的盐酸金霉素多抗血清,为盐酸金霉素单抗的制备奠定了基础。
译  名：
Synthesis of Chlortetracyline Artificial Antigen and Preparation of Its Mice Polyclonal Antiserum
作  者：
LI Jing1,2,ZHI Ai-min1,LI Qing-mei1,LIU Qing-tang1,HOU Yu-ze2, HU Xiao-fei1,DENG Rui-guang1,ZHANG Gai-ping1*(1.Key Laboratory for Animal Immunology of the Ministry of Agriculture,Henan Academy of Agricultural Sciences,Zhengzhou 450002,China;2.Food and Bioengineering College,Henan University of Science and Technology,Luoyang 471003,China)
关键词：
chlortetracyline;polyclonal antiserum;artificial antigen
摘  要：
Chlortetracyline(CTC) was conjugated the carrier protein bovine serum albumin(BSA) and ovalbumin(OVA) by the method of EDC to prepare immunizing antigen BSA-CTC and coating antigen OVA-CTC.The antigen BSA-CTC and OVA-CTC were identified by ultra violet scanning spectrum and SDS-PAGE.The results showed that compared with BSA and CTC,UV spectrogram of BSA-CTC changed and indicated that the hapten CTC was linked to carrier proteins.The titers and sensitivity of chlortetracyline polyclonal antibody were determined by indirect ELISA and blocking ELISA,which showed that the indirect ELISA titers against CTC of five immunized BALB/c mice were above 1×10-3 and the IC50 of No.3 mouse was 71.47 ng/mL.The high-titer and high-specific chlortetracyline polyclonal antibody was got,which made a foundation to prepare monoclonal antibody of chlortetracyline.

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