[恶性疟原虫红细胞结合抗原175单克隆抗体的制备与鉴定]

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

作者信息

Hao Wen-bo, Hong Yan-hua, Sun Xiao-min, Ning Yunshan, Zhang Dong-mei, Pan Wei-qing, Li Ming

机构信息

School of Biotechnology, Southern Medical University, Guangzhou 510515, China.

出版信息

Di Yi Jun Yi Da Xue Xue Bao. 2005 Sep;25(9):1169-71.

PMID:16174591

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.

摘要

目的

制备并鉴定抗恶性疟原虫红细胞结合抗原175（EBA-175）的单克隆抗体（mAb）。

方法

用纯化的重组EBA-175免疫BALB/c小鼠，采用杂交瘤技术制备抗EBA-175的mAb。采用酶联免疫吸附测定（ELISA）和蛋白质印迹法对mAb进行鉴定。

结果

获得了6株抗EBA-175抗原的单克隆抗体，其中5株鉴定为IgG1，1株鉴定为IgG2a。这些单克隆抗体在腹水中的效价为1:12,800至1:25,600，在上清液中的效价为1:256至1:512，ELISA显示4株单克隆抗体（1F3、2H5、4A1和4H9）与恶性疟原虫有特异性结合。蛋白质印迹法显示其中3株单克隆抗体可识别EBA-175蛋白。