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采用恶性疟原虫重组富组氨酸蛋白 (HRP- )融合表达蛋白免疫 BAL B/ c小鼠 ,取脾细胞与 SP2 / 0细胞融合 ,经 3次 EL ISA筛选 ,共获得 6株分泌抗恶性疟原虫重组 HRP- 单克隆抗体的杂交瘤细胞株 (1B11、1D10、2 E7、3A3、3F9、4G5 ) ,用有限稀释法进行克隆、亚克隆及扩大培养 ,并用杂交瘤细胞经腹腔接种 BAL B/ c小鼠制备腹水。 6株杂交瘤细胞分泌的单克隆抗体 (Mc Ab)经琼脂双扩散鉴定均为 Ig G1亚类 ,Dot- EL ISA及 Western blot分析显示 6株 Mc Ab都能与重组 HRP- 抗原发生特异性反应 ,但其中只有 2 E7和 3A3在 Dipstick免疫胶体金反应中能与恶性疟原虫培养上清中的天然 HRP- 结合
BALB / c mice were immunized with recombinant protein of Plasmodium falciparum-rich histidine-rich protein (HRP-) and spleen cells were fused with SP2 / 0 cells. After 3 times of EL ISA screening, 6 strains of BALB / c mice (1B11, 1D10, 2E7, 3A3, 3F9, and 4G5) of protozoal recombinant HRP-monoclonal antibody were cloned, subcloned and expanded by limiting dilution method. The BALB / c mice to prepare ascites. The monoclonal antibodies secreted by 6 hybridoma cells (Mc Ab) were both Ig G1 subclass identified by agar diffusion assay. Dot-ELISA and Western blot analysis showed that all 6 Mc Abs reacted specifically with recombinant HRP-antigen , But only two of them, E7 and 3A3, were able to bind to native HRP- in P. falciparum culture supernatant in the Dipstick immunogold reaction