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目的制备抗恶性疟原虫单克隆抗体，为疟疾诊断及疟疾疫苗等方面的研究提供良好的材料。方法以恶性疟原虫复合重组融合蛋白为免疫原，经细胞融合及ＥＬＩＳＡ方法筛选获得阳性杂交瘤细胞。对其中３株２Ｈ１２、３Ｄ５、４Ｅ５进行了染色体核型分析，对其分泌的单克隆抗体作ＳＤＳ－ＰＡＧＥ、Ｗｅｓｔｅｒｎｂｌｏｔ及Ｄｏｔ－ＥＬＩＳＡ鉴定，并初步观察了３株单抗对恶性疟原虫（海南分离株）的体外抑制作用。结果两次细胞融合共获得８株阳性克隆，其中３株即２Ｈ１２、３Ｄ５、４Ｅ５的ＯＤ值超过阳性对照，传至２０代以上经ＥＬＩＳＡ检测仍呈强阳性反应，将其腹腔接种给ＢＡＬＢ／ｃ小鼠获得血性腹水，抗体滴度为１∶１２８００。Ｗｅｓｔｅｒｎｂｌｏｔ及Ｄｏｔ－ＥＬＩＳＡ分析显示３株单抗均与相应蛋白发生特异性反应。抑制试验结果表明：３株单抗对恶性疟原虫的体外生长和发育均有一定的抑制作用，抑制率分别为４９％±３％、７６％±６％、４１％±２％。结论已获得稳定分泌抗恶性疟原虫复合重组抗原单克隆抗体的杂交瘤细胞株。
Objective To prepare monoclonal antibodies against Plasmodium falciparum and provide a good material for the research of malaria diagnosis and malaria vaccine. Methods Plasmodium falciparum recombinant fusion protein was used as immunogen. The positive hybridoma cells were obtained by cell fusion and ELISA screening. Three karyotypes of 2H12, 3D5 and 4E5 were analyzed for their karyotypes. The secreted monoclonal antibodies were identified by SDS-PAGE, Western blot and Dot-ELISA. Three McAbs against P. falciparum (Hainan isolate Strain) in vitro inhibition. Results Eight positive clones were obtained by two cell fusion. The OD value of three strains, 2H12, 3D5 and 4E5, surpassed that of the positive control and reached to be more than 20 passages. The results of ELISA showed that there was still a strong positive reaction. The BALB / c Mice received bloody ascites, antibody titers of 1: 12800. Western blot and Dot-ELISA analysis showed that the three McAbs reacted specifically with the corresponding proteins. The results of inhibition test showed that the three McAbs could inhibit the growth and development of Plasmodium falciparum in vitro with the inhibitory rates of 49% ± 3%, 76% ± 6% and 41% ± 2%, respectively. Conclusion Hybridoma cell lines stably secreting monoclonal antibodies against Plasmodium falciparum complexed recombinant antigen have been obtained.