小鼠IFIT1多克隆抗体的制备  

Preparation of polyclonal antibody of mouse IFIT1

作　　者：郭晓姝[1,2] 李洪涛[1] 方海立[2] 王军平[1] 粟永萍[1] 

机构地区：[1]第三军医大学军事预防医学院防原医学教研室,全军复合伤研究所,创伤,烧伤与复合伤国家重点实验室,重庆4000382 [2]重庆医科大学基础医学院生理学教研室,重庆400016

出　　处：《第三军医大学学报》2007年第15期1462-1464,共3页Journal of Third Military Medical University

基　　金：国家重点基础研究发展规划("973"项目)(G1999054201);国家自然科学基金海外学者合作研究项目(30328025);创伤;烧伤与复合伤国家重点实验室开放课题(2004)~~

摘　　要：目的获取小鼠IFIT1(Interferon-induced protein with tetratricopetide repeats 1)特异性多克隆抗体。方法扩增培养高效表达IFIT1的重组子pMAL-C2X-IFIT1,超声破碎后的细胞上清过Amylose Pre-packed column亲和层析柱,将所得纯化的融合蛋白MBP-IFIT1作为抗原,添加福氏佐机剂后免疫兔,收集抗血清,用Western blot和ELISA方法测定抗血清特异性反应和效价。结果纯化的MBP-IFIT1可诱导兔产生特异性免疫应答,所得抗体能够特异性识别融合蛋白中的IFIT1,ELISA结果显示其效价为1∶12800。结论MBP-IFIT1具有良好的抗原性,以该融合蛋白为抗原免疫兔成功制备出IFIT1特异性多克隆抗体。Objective To acquire specific polyclonal antibody of mouse IFIT1 （ Interferon-induced protein with tetratricopetide repeats 1 ）. Methods The single clone of pMAL-C2X-IFIT1, confirmed previously to highly express MBP-IFIT1 fusion protein, was cultivated and amplified. After ultrasonic fragmentation and centrifugation, the MBP-IFIT1 fusion protein was purified from the supernatant by using the Amylose Pre-packed column. Then, the MBP-IFIT1 fusion protein, mixed with Freund＇ s adjuvant, was injected into rabbits twice with two-week interval. The rabbit serum was collected its anti-IFIT1 specific reaction was detected by Western , then its anti-IFIT1 titer was detected by ELISA, and blot. Results The rabbit serum immunized with purifled MBP-IFIT1 protein showed specific anti-IFIT1 ability, and its antibody titer was about 1：12 800. Conclusion The purified MBP-IFIT1 fusion protein possesses favorable antigenicity, and highly specific polyclonal antibody of mouse IFIT1 can be obtained by using purified MBP-IFIT1 as antigen to immunize rabbit.

关 键 词：IFIT1 多克隆抗体 融合蛋白 抗原性 免疫应答 

分 类 号：R392-33[医药卫生—免疫学] R392.11[医药卫生—基础医学]