重组人骨形态发生蛋白-7抗体的制备及鉴定

目的利用重组人骨形态发生蛋白-7（rhBMP-7）制备抗体,为BMP-7临床应用奠定基础。方法采用皮内多点注射法将rhBMP-7免疫新西兰大白兔4次,每次间隔2周。免疫后颈动脉插管取血。ELISA法测定抗血清效价,蛋白免疫印迹技术确定抗血清特异性、最佳稀释度、灵敏度。结果抗血清效价为1：640000;通过从小鼠骨组织及肾脏组织中提取的BMP-7证实抗体有很好的特异性,抗血清的最佳稀释度为1：5000-1：50000,灵敏度为1～100ng。结论成功制备了rhBMP-7抗体,该抗体具有很高的特异性及灵敏度。

Preparation and identification of recombinant human bone morphogenetic protein-7's antibody

Objective To prepare the recombinant human bone morphogenetic protein-7（rhBMP-7） ＇s antibody, and lay the foundation for the clinical application of BMP-7. Methods New Zealand white rabbits were immuned 4 times by rhBMP-7 in the way of subcutaneous multi-point injection,with the time interval of 2 weeks. Then the blood samples were obtained through carotid artery intubation ,and the valence of antiserum was detected by using the method of ELISA , the antiserum＇s specificity, the best degree of dilution and sensitivity were measured by West blotting. Results The valence of rhBMP-7＇s antiserum was 1 ： 640 000. The BMP-7 which extracted from the bone and kidney of mouse showed that the antiserum had high specificity. The best degree of dilution was 1：5 000 to 1：50 000, while the sensitivity was 1 to 100 ng. Conclusions The antibody of rhBMP can be prepared successfully, which possess high specificity and sensitivity.