摘要
目的:制备高效价E血清型沙眼衣原体(CT)的多克隆抗体。方法:用hep-2细胞培养制备并用超速离心方法纯化E血清型CT作为抗原,采用弗氏佐剂乳化抗原,背部皮下多点免疫4只家兔,收获免疫后血清制备多克隆抗体,用Western blot方法进行鉴定,ELISA法检测抗体水平。结果:所有家兔在全程免疫后都产生了高效价的血清抗体,最高滴度达1:1600。Western blot实验结果显示,本研究制备的CT血清抗体能与CT全菌体抗原发生特异结合。结论:CT菌体抗原免疫家兔可成功制备高效价的血清抗体,为建立沙眼衣原体免疫检测方法打下了基础。
Objevtive：To prepare polyclonal antibody against Chlamydia trachomatis （CT） serovar E.Methods： Chlamydia trachomatis was cultivated and proliferated in Hep-2 cell culture system with RPMI 1640 culture medium. Thereafter, the purified CT was harvested using supercentrifuge method. Frepare CT antigen containing Freund adjuvant by routine method. Four rabbits were immunized with the prepared antigen by subcutaneous injection in various sites on back. The sera of these immunized rabbits were collected for preparation of polyclonal antibody. Immune response was identified by Western blot in order to determine its titer by ELISA. Results： All of the immunized rabbits produced high-titer serum antibody after the second booster. The highest titer of antibody against CT was 1：1600. Western blot showed the antisbody can bind to the CT protein specifically. Conclusion： The polyclonal antibody against CT, with high titer, is successfully prepared. It laid a foundation of development of method for detection of CT.
出处
《温州医学院学报》
CAS
2007年第4期344-346,共3页
Journal of Wenzhou Medical College
基金
浙江省医药卫生重点科技项目科研基金资助项目(2005ZD011)
温州市科技局科研基金资助项目(Y2004A007)。
关键词
沙眼衣原体
抗体
免疫性
Chlamydia trachomatis
antibody
immunity
