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作 者:安新玲[1] 王志宇[1] 刘肖帅[1] 葛鲁娜[1] 朱有名[1] 韩金祥[1] 王世立[1]
机构地区:[1]山东省医药生物技术研究中心国家卫生部生物技术药物重点实验室,济南250062
出 处:《山东医药》2009年第43期27-28,共2页Shandong Medical Journal
基 金:国家863计划资助项目(2003AA223532)
摘 要: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。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.
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