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目的:利用重组狂犬病病毒糖蛋白(RVG)免疫人源Ig M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Ig M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Ig M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。
OBJECTIVE: To immunize human Ig M transgenic mice with recombinant rabies virus glycoprotein (RVG) to prepare a monoclonal antibody against human recombinant RVG protein. The immunological characteristics of the monoclonal antibody were preliminarily identified. Methods: Human IgM transgenic mice were immunized with recombinant RVG protein as antigen and hybridoma cell lines were screened by hybridoma technique. Human double antibody sandwich ELISA was used to identify the human monoclonal antibodies and their antibody types , And its specificity and ability to bind with inactivated rabies virus CVS-11 strain were identified. Results: Five hybridoma cell lines stably secreting anti-recombinant RVG were established and named as 5D1, 6H11, 9A3, 15D6 and 19E6 respectively, all of which were human Ig M immunoglobulins. All the 5 monoclonal antibodies Which can specifically recognize recombinant RVG protein. Three of them can specifically bind to the inactivated rabies virus CVS-11 strain. Conclusion: Monoclonal antibody against specific recombinant human RVG protein was screened and prepared. It can specifically bind to the inactivated rabies virus (CVS-11) strain and lay the foundation for the further development of anti-rabies antibody.