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[目的 ]制备抗恶性疟原虫乳酸脱氢酶 (LDHp)单克隆抗体 (McAb) ,并对其特异性进行鉴定。[方法 ]用纯化的LDHp重组抗原免疫BALB/c小鼠 ,采用杂交瘤技术制备McAb ,筛选分泌高滴度McAb的杂交瘤细胞株 ,测定其免疫球蛋白亚类及其效价 ,ELISA、Westernblot试验分析其特异性。 [结果 ]筛选出 2A5和1H10两株能稳定分泌抗LDHpMcAb的杂交瘤细胞株 ,两株单抗均为IgG2b,2A5和 1H10培养上清的ELISA效价分别为 1∶5 12和 1∶2 5 6 ,腹水效价分别为 1∶2 5 6 0 0和 1∶12 80 0 ,两株单抗与间日疟原虫、红细胞、弓形虫、日本血吸虫等抗原均不发生交叉反应 ,能识别恶性疟原虫 33kDa的虫源蛋白。 [结论 ]制备的抗LDHp杂交瘤细胞株能分泌高滴度和高特异性的单抗
[Objective] The research aimed to prepare the monoclonal antibody against Plasmodium falciparum lactate dehydrogenase (LDHp) and identify its specificity. [Method] BALB / c mice were immunized with purified LDHp recombinant antigens and McAbs were prepared by hybridoma technique. The hybridoma cell lines secreting high titer McAbs were screened and their immunoglobulin subclasses and their titer were determined. ELISA, Western blot Experimental analysis of its specificity. [Results] The two McAbs 2A5 and 1H10 that could stably secrete anti-LDHpMcAb were screened out. The two monoclonal antibodies were both IgG2b. The ELISA titers of 2A5 and 1H10 culture supernatant were 1:5 12 and 1: 25, respectively 6, ascites titers were 1:25 600 and 1:12 80 0, respectively. The two McAbs did not cross-react with antigens such as Plasmodium vivax, erythrocytes, Toxoplasma gondii and Schistosoma japonicum, Protozoa 33kDa insect protein. [Conclusion] The prepared anti-LDHp hybridoma cell lines can secrete monoclonal antibody with high titer and high specificity