His6免疫原的制备及免疫小鼠抗体效价检测  

Preparation of Artificial Immunogen of His6 Peptides and Its Antibody Detection in Mice

作　　者：朱彦彩[1] 王选年[1] 银梅[2] 乔新安[1] 吴艳云[1] 熊静[2] 

机构地区：[1]河南农业大学牧医工程学院,河南郑州450002 [2]河南科技学院动物科学学院,河南新乡453003

出　　处：《河南农业科学》2007年第12期116-119,共4页Journal of Henan Agricultural Sciences

摘　　要：采用EDC一步法将His6多肽分别与匙孔血蓝蛋白(KLH)和牛血清白蛋白(BSA)偶联,制备两种人工结合抗原KLH-His6和BSA-His6。将制备的KLH-His6作为免疫原,皮下分5点注射免疫6只健康6周龄雌性Balb/c小鼠;首次免疫取KLH-His6(50μg/只)与氟氏完全佐剂(FCA)充分乳化,每只免疫200μL,以后每间隔两周,同法进行第2,3次免疫,用氟氏不完全佐剂(FIA)乳化。每次免疫8 d后尾部采血,以BSA-His6为包被抗原检测抗His6抗体。结果表明,成功制备了His6免疫原,并免疫了小鼠;间接ELISA检测不同时期小鼠抗体效价最高达到1∶8 000,免疫小鼠的抗体效价水平整体呈上升趋势,为His6单隆抗体的制备奠定了基础。His6 polypeptide was coupled with keyhole limpet hemocyanin （KLH） and bovine serum albumin （BSA） using EDC method, from which two artificial antigens, KLH--His6 and BSA --His6 were developed. Then KLH--His6 was used as immunogen to immunize six healthy female Balb/c mice of six weeks by five points of subcutaneous injection. After the first time of immunization,50μg of KLH--His6 from each mouse was taken and was emulsified with freund＇s complete adjuvant （FCA）. Afterward, the second and the third immunization were carried out in the same way as before at an interval of two weeks, and the same concentration of KLH--His6 was emulsified with freund＇s incomplete adjuvant （FIA）. After 8 days of each immunization, blood samples were collectd from the mouse tails. The antibody to His6 was detected by indirect ELISA used BSA--His6 as coating antigen. The results showed that the immunogen His6 was developed successfully, which could immunize mice. Indirect ELISA method is used to detect different periods of the immunized mice antibody and the highest titer can achieve 1 ： 8 000. The antibody titer of immunized mice was keeping an increasing tendency. The experiment results laid the foundation for preparation of His6 monoclonal antibody.

关 键 词：His6多肽 免疫原制备 间接ELISA 抗体效价 

分 类 号：S852.42[农业科学—基础兽医学]