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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 anti-human cardiac troponin I antibody and identify it. Methods New Zealand rabbits were immunized with synthetic cTnI protein to prepare polyclonal antibodies. The antiserum titer and specificity were determined by ELISA and Western blot. Results The titer of antiserum ELISA was 1:16 000 and the sensitivity of cTnI was 10 μg / L. Western blot showed that the antibody can effectively detect the expression of cTnI protein in the prokaryotic cells and rat heart tissue extracts. Conclusion The polyclonal antibody of human cTnI was successfully prepared by this method, which laid the foundation for further study on the structure and function of cTnI and provided a reference for efficient preparation of highly specific monoclonal antibodies.