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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。
To prepare McAbs of Plasmodium falciparum lactate dehydrogenase (LDHp) and identify their specificity, BALB / c mice were immunized with recombinant LDHp recombinant antigen and McAbs were prepared by hybridoma technique. Hybridoma cell lines secreting high titer McAbs were tested for their immunoglobulin subclasses and their potency. ELISA and Westen blotting assays were used to analyze their special conductivity. As a result, two hybridoma cell lines, 2A5 and 1H10, capable of stably secreting anti-LDHp McAb were prepared, and the two McAbs were both IgG2b. ELISA titers of 2A5 and 1H10 culture supernatants were 1: 512 and 1: 256, respectively, The titer was 1: 25600 and 1: 12800, respectively. The two McAbs did not cross-react with P. vivax, erythrocytes, Toxoplasma gondii, Schistosoma japonicum and other antigens and could recognize the 33KDa protein of P. falciparum. It is proved that the prepared anti-LDHp hybridoma cell lines can secrete high titers and high specific monoclonal antibodies.