全人源抗狂犬病病毒糖蛋白抗体的制备及鉴定

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?

Generation and identification of fully human monoclonal antibodies against the rabies virus glycoprotein

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.