【摘要】 为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,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,筛选出分泌高滴度McAb的杂交瘤细胞株,测定其免疫球蛋白亚类及其效价,ELISA、Westen blotting试验分析其特导性。结果,制备出2A5和 1H10两株能稳定分泌抗LDHp　McAb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1:512和1:256,腹水效价分别为1:25600和1:12800,两株单抗与间日疟、红细胞、弓形虫、日本血吸虫等抗原均不发生交叉反应,能识别恶性疟原虫的33Kda虫源蛋白。证明制备的抗LDHp杂交瘤细胞株能分泌高滴度和高特异性的单抗。更多还原

【Abstract】 To prepare and characterize the monoclonal antibodies (McAbs) against lactate dehydrogenase of the Plasmodium falciparum (LDHp). Methods: BALB/c mice were immunized with purified recombinant LDHp and McAbs against LDHp were prepared by using hybridoma technique. The characterization of the McAbs was performed by ELISA and Western-blotting test. Result: Two McAbs cell lines against LDHp antigen were obtained. The immunoglobulin class/ subclasses of two McAbs were both IgG2b. The titer of two McAbs (2A5, 1H10) in the ascites was 1 = 25600 and 1: 12800, and that in the supernatant were 1:512 and 1 :256 respectively. The result of ELBA indicated that 2 McAbs only reacted with Plasmodium falciparum, but not with normal human erythrocytes, Plasmodium vivax, Toxoplasma gondii or Schistosoma japonicam. The 2 McAbs could recogniz 33KDa protein from Plasmodium falciparum that was defined as LDHp by Western-blotting test. Conclusion: Two hybridoma cell lines against LDHp secreting higt titer of mAbs with high specificity were established.更多还原