兔抗人心肌肌钙蛋白Ⅰ抗体的制备与鉴定  

Preparation and Characterization of Rabbit Antibody Against Human Cardiac Troponin I

作　　者：杨能[1] 徐霞[1] 谢闺娥[1] 

机构地区：[1]广州医学院检验系,广州510182

出　　处：《热带医学杂志》2006年第1期20-22,共3页Journal of Tropical Medicine

基　　金：广东省医学科研基金(No.A2000265)。

摘　　要：目的制备兔抗人心肌肌钙蛋白I抗体并进行鉴定。方法用人工合成的cTnI蛋白免疫新西兰兔制备多克隆抗体,ELISA及Western blot方法检测制备的抗血清效价和特异性。结果此次获得的抗血清 ELISA方法检测抗体滴度达到了1:16 000,检测cTnI的灵敏度最低为10μg/L。Western blot检测表明该抗体可有效识别原核细胞表达及大鼠心脏组织提取物中存在的cTnI蛋白。结论应用该方法成功制备了人cTnI的多克隆抗体,为进一步研究cTnI的结构与功能打下基础,并为有效制备高特异性的单克隆抗体提供借鉴。Objective To prepare rabbit antibody against human cardiac troponin I by using recombinant cTnI protein. Methods New Zealand rabbit was immunized with the E. coli expressed and purified rcombinant cTnI protein. Polyclonal antiserum was analyzed by ELISA （enzyme linked immunsosrbent assay） and Western blot. Results The titer of the antiserum obtained in this experiment determined by ELISA was up to 1.16 000 and the sensitivity was at least 10 μg/L. Western blot analysis assay confirmed that the antibodies could detect both prokaryotic expressed cTnI, and the native cTnI in the cardiac tissue of mouse. Conclusion Polyclonal anti-cTnI antibodies with high specificity and affinity were prepared by using purified recombinant cTnI as immunogen. This polyclonal antibodies preparation lays the foundation for further research on the structure and function of cTnI, as well as serving as an useful reference for the preparation of monoclonal antibody.

关 键 词：CTNI 多克隆抗体 兔 

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