抗恶性疟McAb的筛选与恶性疟免疫诊断研究

用单克隆抗体(McAb)夹心斑点免疫金银染色法(Dot-IGSSA)对11株抗恶性疟McAbs和4株抗食蟹猴疟原虫McAbs进行了筛选,其中McAbs11G5、13A2、13A1可包被到硝酸纤维素膜(NcF)上作为捕获抗体,McAb14D9可用于胶体金探针的标记。此外,为增加胶体金的标记效果和胶体金标记McAb14D9探针的敏感性,还对McAb14D9的等电点(pI)进行了测定,其pI为6.35。同时用筛选出的最佳McAbs对10份恶性疟病人感染血和10份健康人血样品进行了检测,并对这几株McAbs的交叉反应性进行了测定,结果表明McAbs11G5、13A1与14D9夹心时,对云南株和安微株恶性疟原虫红内期抗原呈现交叉反应,而且McAb13A1与14D9夹心时还可以用于检测间日疟抗原。

The screening of monoclonal antibodies against Plasmodium falciparum and study on the immunodiagnosis of Plasmodium falciparum

11 strains of monoclonal antibodies ( McAbs ) against Plasmodium faliciparum and 4 strains of McAbs against Plasmodium cynomolgi was screened by McAb sandwich dot-immunogold-silver staining assay ( Dot-IGSSA ) . Of which McAbs 11G5, 13A2 and 13Al could be coated to nitrocelluose filter ( NCF ) as capturing ntibodies and McAb 14D9 could be labeled with colloidal gold. In addition, in order to increase the labeled efficiency of colloidal gold and the sensitivity of McAb 14D9 labeled with colloidal gold, the isoelectric point ( pI ) of McAb 14D9 which were used for labelling was measured by isoelectric focusing in polyacrylamide gel. The pI of McAb 14D9 was 6.35. At the same time, infected blood samples from 10 patients with Plasmodium falciparum and normal blood samples from 10 healthy persons were examined by McAb sandwich Dot-IGSSA using optimal McAbs of screening and the cross-reactions of these McAbs with different species and isolates of malaria parasites were also detected by McAb sandwich Dot-IGSSA. The results showed that the McAbs llG5,13Al and 14D9 could be used to examine the antigens of asexual blood stages of Plasmodium falciparum of isolats from Yunan and Anhui, China, however, McAb 13Al and 14D9 also cross-reacted with Plasmodium vivax.