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重组人骨形态发生蛋白-7抗体的制备及鉴定

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

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【作者】 安新玲; 王志宇; 刘肖帅; 葛鲁娜; 朱有名; 韩金祥; 王世立;

【Author】 AN Xin-ling,WANG Zhi-yu,LIU Xiao-shuai,GE Lu-na,ZHU You-ming,HAN Jin-xiang,WANG Shi-li(Shangdong Medicinal Biotechnology Center,Key Laboratory for Biotech-Drugs of Ministry of Health,Jinan 250062,P.R.China)

【机构】 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;

【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∶640 000;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∶5 000~1∶50 000,灵敏度为1~100 ng。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。

【Abstract】 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.

【关键词】 骨形态发生蛋白-7; 抗体; 效价;
【Key words】 bone morphogenetic protein-7; antibody; vlence;
【基金】 国家863计划资助项目(2003AA223532)
  • 【文献出处】 山东医药 ,Shandong Medical Journal , 编辑部邮箱 ,2009年43期
  • 【分类号】R392.1
  • 【被引频次】1
  • 【下载频次】41
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