[恶性疟原虫PfCP-2.9嵌合蛋白特异性单克隆抗体的制备与鉴定]
[Preparation and characterization of monoclonal antibody specific to PfCP-2.9 chimeric protein of Plasmodium falciparum].
作者信息
Wang Rui, Qian Feng, Qu Li, Pan Wei-qing
机构信息
Department of Etiologic Biology, the Second Military Medical University, Shanghai, China.
出版信息
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2006 Aug;24(4):247-50.
OBJECTIVE
To prepare and characterize monoclonal antibody against a malaria vaccine candidate, PfCP-2.9 chimeric protein of Plasmodium falciparum.
METHODS
BALB/c mice were immunized with PfCP-2.9, and the spleen cells were used for fusion with SP2/0 cells. The monoclonal antibodies were analyzed by ELISA, Western blotting as well as growth inhibition assay.
RESULT
A monoclonal antibody was obtained. It interacted with the PfCP-2.9 recombinant protein by ELISA and Western blotting. The interaction of the monoclonal antibody with the protein was reduction-sensitive, indicating that the antibody recognized a conformational epitope. Moreover, the antibody also recognized the cultured parasites of P. falciparum by indirect immunofluorescent antibody test (IFA). When tested by growth inhibition assay, the antibody significantly inhibited parasite growth in vitro of 56% inhibition rate at the antibody concentration of 0.3 mg/ml.
CONCLUSION
A monoclonal antibody against PfCP-2.9 malaria vaccine candidate has been obtained, which recognizes a conformational epitope of the protein and natural protein.
目的
制备并鉴定针对疟原虫疫苗候选物恶性疟原虫PfCP-2.9嵌合蛋白的单克隆抗体。
方法
用PfCP-2.9免疫BALB/c小鼠，取其脾细胞与SP2/0细胞进行融合。通过酶联免疫吸附测定（ELISA）、蛋白质印迹法以及生长抑制试验对单克隆抗体进行分析。
结果
获得了一种单克隆抗体。通过ELISA和蛋白质印迹法，该抗体与PfCP-2.9重组蛋白发生相互作用。该单克隆抗体与该蛋白的相互作用对还原敏感，表明该抗体识别一个构象表位。此外，通过间接免疫荧光抗体试验（IFA），该抗体也识别恶性疟原虫的培养寄生虫。在生长抑制试验中检测时，在抗体浓度为0.3 mg/ml时，该抗体显著抑制体外寄生虫生长，抑制率达56%。
结论
已获得一种针对疟原虫疫苗候选物PfCP-2.9的单克隆抗体，该抗体识别该蛋白的一个构象表位以及天然蛋白。
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