文章摘要
王晓蕾,朱 进,哈 卓,张晓萍,杨 岚,杨晓明,刘云成,Mason Lu,冯振卿.全人源抗狂犬病病毒糖蛋白抗体的制备及鉴定[J].南京医科大学学报,2017,(2):160~164
全人源抗狂犬病病毒糖蛋白抗体的制备及鉴定
Generation and identification of fully human monoclonal antibodies against the rabies virus glycoprotein
投稿时间:2016-04-13  
DOI:10.7655/NYDXBNS20170205
中文关键词: 人源IgM转基因小鼠  狂犬病病毒糖蛋白  全人源单克隆抗体
英文关键词: humanized IgM transgenic mice  rabies virus glycoprotein(RVG)  human monoclonal antibody
基金项目:江苏省社会发展项目(BE2011842);国家自然科学基金(81273325)
作者单位
王晓蕾 南京医科大学病理学系,卫生部抗体技术重点实验室,江苏 南京 210029
南京医科大学附属南京医院病理科,江苏 南京 210006 
朱 进 卫生部抗体技术重点实验室,江苏 南京 210029
南京军区军事医学研究所,江苏 南京 210002 
哈 卓 东竺明生物技术有限公司,黑龙江 大庆 163316
福瑞邦生物科技有限公司,黑龙江 大庆 163316 
张晓萍 东竺明生物技术有限公司,黑龙江 大庆 163316
福瑞邦生物科技有限公司,黑龙江 大庆 163316 
杨 岚 东竺明生物技术有限公司,黑龙江 大庆 163316
福瑞邦生物科技有限公司,黑龙江 大庆 163316 
杨晓明 东竺明生物技术有限公司,黑龙江 大庆 163316
福瑞邦生物科技有限公司,黑龙江 大庆 163316 
刘云成 福瑞邦生物科技有限公司,黑龙江 大庆 163316 
Mason Lu 东竺明生物技术有限公司,黑龙江 大庆 163316
University of Texas MD Anderson Cancer Center,Houston,Texas 77030 
冯振卿 南京医科大学病理学系,卫生部抗体技术重点实验室,江苏 南京 210029 
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中文摘要:
      目的:利用重组狂犬病病毒糖蛋白(RVG)免疫人源IgM转基因小鼠,制备全人源抗重组RVG蛋白单克隆抗体,并对其免疫学特性进行初步鉴定。方法:以重组RVG蛋白作为抗原免疫人源IgM转基因小鼠,采用杂交瘤技术制备筛选全人源抗重组RVG蛋白杂交瘤细胞株,双抗体夹心ELISA实验鉴定单抗的人源性及抗体类型,并对其特异性及与灭活狂犬病病毒CVS-11株的结合能力进行鉴定。结果:建立了5株稳定分泌抗重组RVG的全人源单克隆抗体杂交瘤细胞株,分别命名为5D1?6H11?9A3?15D6?19E6,均为人源IgM免疫球蛋白,5株单抗均能特异性识别重组RVG蛋白,其中3株能与灭活狂犬病病毒CVS-11株特异性结合。结论:筛选制备了特异性全人源抗重组RVG蛋白的单克隆抗体,能与灭活狂犬病病毒CVS-11株特异性结合,为进一步研制用于狂犬病防治的抗体药物奠定了基础。
英文摘要:
      Objective:To generate human monoclonal antibodies(mAbs)against the rabies virus glycoprotein(RVG)by immunizing the mice carrying human immunoglobulin transloci, and further to identify their characters. Methods:The human IgM transgenic mice were immunized with the rabies virus glycoprotein. The human anti-RVG hybridoma cell lines were screened and produced by using the classical hybridoma technique. The specificity and isotype of mAbs were determined by the double antibody sandwich enzyme- linked immunosorbent assay(ELISA). Also,the binding properties with the inactivated rabies virus CVS-11 were analyzed. Results:Five hybridoma cell lines were established,they can stably produce human anti-rabies virus IgM monoclonal antibodies(5D1,6H11,9A3,15D6,and 19E6). These mAbs specifically recognized the r-RVG and three human anti-r-RVG mAbs specifically combined with the inactivated rabies virus CVS-11 strain. Conclusion:Hybridoma cell lines were successfully established,and can stably produce human anti-rabies virus IgM monoclonal antibodies. Human anti-r-RVG mAbs could specifically combine with the inactivated rabies virus CVS-11 strain. This may be a foundation for further development of vaccine to prevent and control rabies.
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