南方医科大学学报

南方医科大学学报 ›› 2005, Vol. 25 ›› Issue (09): 1169-1171.

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抗恶性疟原虫EBA-175单克隆抗体的制备与初步鉴定

郝文波1, 洪艳华1, 孙晓敏1, 宁云山1, 张冬梅2, 潘卫庆2, 李明1   

  1. 1. 南方医科大学生物技术学院, 广东, 广州, 510515;
    2. 第二军医大学病原生物学教研室, 上海, 200433
  • 出版日期:2005-09-20 发布日期:2005-09-20
  • 基金资助:
    收稿日期:2005-7-12。
    基金项目:国家自然科学基金(30170839)
    作者简介:郝文波,女,讲师,电话:020-61648321,E-mail:haowa@126.com
    通讯作者:李明,教授,博士生导师,电话:020-61648550,E-mail:min-gli@fim m u.com.

Preparation and characterization of monoclonal antibodies against erythrocyte-binding antigen 175 of Plasmodium falciparum

HAO Wen-bo1, HONG Yan-hua1, SUN Xiao-min1, NING Yunshan1, ZHANG Dong-mei2, PAN Wei-qing2, LI Ming1   

  1. 1. 南方医科大学生物技术学院, 广东, 广州, 510515;
    2. 第二军医大学病原生物学教研室, 上海, 200433
  • Online:2005-09-20 Published:2005-09-20

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摘要： 目的 制备抗恶性疟原虫EBA-175的单克隆抗体(McAb),为恶性疟原虫的诊断及疫苗研究奠定基础。方法 以恶性疟原虫EBA-175(II区F2段)重组抗原免疫BALB/c小鼠,采用杂交瘤技术制备McAb,间接ELISA筛选分泌高滴度McAb的杂交瘤细胞株,测定其免疫球蛋白亚类及其效价,ELISA、Westernblot试验分析其特异性。结果 筛选获得6株稳定分泌抗重组EBA-175McAb的杂交瘤细胞株1F3、2E8、2H5、4A1、4C3、4H9,6株McAb经鉴定其亚类5株为IgG1,1株为IgG2a,6株McAb的培养上清ELISA效价为1:256~1:512,腹水效价为1:12800~1:25600,间接ELISA显示其中4株McAb1F3、2H5、4A1、4H9能与恶性疟原虫抗原发生特异性结合,而只有1F3、4A1、4H9在Westernblot试验中识别恶性疟原虫Mr约35000的虫源蛋白。结论 获得了能稳定分泌高特异性抗EBA175McAb的杂交瘤细胞。

Abstract: Objective To prepare and characterize the monoclonal antibodies (mAbs) against erythrocyte-binding antigen 175 of Plasmodium falciparum (EBA-175). Methods BALB/c mice were immunized with purified recombinant EBA-175 and mAbs against EBA-175 were prepared by means of hybridoma technique. Enzyme-linked immunosorbent assay (ELISA) and Western blotting were employed for characterization of the mAbs. Results Six McAbs against EBA-175 antigen were obtained, 5 of which were identified as IgG1 and one as IgG2a. The titer of these aAbs was 1:12 800 to 1:25 600 in the ascites and 1:256 to 1:512 in supernatant, and ELISA demonstrated specific binding of the 4 mAbs (1F3, 2H5, 4A1 and 4H9)with Plasmodium falciparum. Three of these mAbs recognized the protein of EBA-175 as shown by Western blotting. Conclusion Six hybridoma cell lines secreting the mAbs against EBA175 with high specificity are successfully established.

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