Identification of Recombinant Aldolase of Plasmodium falciparum and its Monoclonal Antibody Preparation
- VernacularTitle:重组恶性疟原虫醛缩酶鉴定及其单克隆抗体的制备
- Author:
Ruijuan ZHANG
;
Huaimin ZHU
;
Hui ZHENG
;
Beifang NING
- Publication Type:Journal Article
- Keywords:
Plasmodium falciparum;
6-bisphosphate aldolase;
Gene expression;
Monoclonal antibody
- From:
Chinese Journal of Parasitology and Parasitic Diseases
1987;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To identify the recombinant aldolase (ALD) of Plasmodium falciparum, and to develop monoclonal antibodies (McAbs) against the recombinant ALD. Methods ALD gene was amplified by PCR from genomic DNA of FCC1/HN strain, and expressed in E.coli DH5?. BALB/c mice were immunized with the recombinant ALD of P. falciparum via celiac injection for 3 times with 2 weeks interval. Three days after a booster injection, spleen cells of the immunized mice were used for producing McAbs. The immune serum was tested by IFAT and Western blotting. Results BALB/c mice immunized with purified aldolase protein developed strong immune response to the antigen, and the titer of specific antibody reached 1∶105 in all immune sera after the third immunization. Moreover, immune sera specifically recognized the cultured P. falciparum. Western blotting showed that the immune sera recognized specifically a Mr 41 000 band of crude malaria antigen. No cross-reaction with human red cells was detected. Seven positive hybridoma cell lines were obtained after 3 rows of selection. All the McAbs′subclasses belong to IgG1. IFAT showed that only 4 McAbs could recognize the cultured P.falciparum. Conclusion Plasmodial aldolase has been successfully expressed and purified, and the established hybridoma cell lines can secrete McAbs specific to the aldolase of P. falciparum.
