抗恶性疟原虫重组谷氨酸脱氢酶单克隆抗体的制备及其鉴定  

PREPARATION AND CHARACTERIZATION OF MONOCLONAL ANTIBODY AGAINST RECOMBINANT GLUTAMATE DAHYDROGENASE OF PLASMODIUM FALCIPARUM

作　　者：李妍[1] 宁云山[1] 李莉[1] 吴锦雅[2] 董文其[1] 李明[1] 

机构地区：[1]第一军医大学热带病教研室,广东广州510515 [2]第一军医大学寄生虫学教研室

出　　处：《中国寄生虫病防治杂志》2004年第3期155-157,共3页Chinese Journal of Parasitic Disease Control

基　　金：广州市科技计划项目 (No .2 0 0 2E3 E40 1 2 )

摘　　要：目的　制备抗恶性疟原虫谷氨酸脱氢酶 (GDH)的单克隆抗体 (McAb) ,并对其特异性进行鉴定。　方法　用柱层析纯化的GDH/GST重组融合蛋白免疫BALB/c小鼠 ,采用杂交瘤技术制备McAb ,用ELISA和有限稀释法筛选出分泌高滴度McAb的杂交瘤细胞株 ,用 protein G亲和层析纯化接种杂交瘤细胞的小鼠腹水 ,测定其免疫球蛋白亚类及其效价 ,Westernblot分析其特异性。　结果　筛选出 6株能稳定分泌抗GDH单克隆抗体的杂交瘤细胞株 ,单抗均为IgG1,Westernblot分析显示 ,6株单抗都与重组的GDH发生特异性结合。　结论　制备的抗GDH杂交瘤细胞株能分泌高滴度和高特异性的单抗 ,为疟疾诊断技术的进一步研究奠定了基础。Objective To prepare monoclonal antibody(McAb) against recombinant glutamate dehydrogenase(GDH) of Plasmodium falciparum (FCC1/HN strain) and identify its characteristics. Methods BALB/c mice were immunized with purified GDH/GST fusion protein. Hybridomas were made using PEG as the fusing agent,and picked out by the methods of ELISA and limited dilution. The McAb was purified by protein-G affinity chromatography. Results Six particular McAbs named 2B9,2G2,3C3,3F5,3F6,5A7 were obtained. The subtype of the six McAbs were all IgG1,with affinity constants (K aff ) of the strains from 1×10 -8 M to 2.8×10 -10 M. Western blot analysis showed that all of the six McAbs strongly and specifically reacted with the recombinant GDH. Conclusion Six hybridomas cell lines secreting high titer of McAbs against GDH of P. falciparum with high specificity are established. This is a basis for further research on the diagnosis of malaria.

关 键 词：疟原虫 恶性 谷氨酸脱氢酶 单克隆抗体 

分 类 号：R382.31[医药卫生—医学寄生虫学]