[Preparation of monoclonal antibodies specific to lactate dehydrogenase of Plasmodium falciparum].
OBJECTIVE
To prepare monoclonal antibodies specific to lactate dehydrogenase of Plasmodium falciparum.


METHODS
The Plasmodium falciparum lactate dehydrogenase (pLDH) gene was amplified from whole blood of malaria patients by PCR and cloned into expression vector pGEX-3X. Recombinant pLDH protein was expressed and purified, and used for immunizing mice to prepare monoclonal antibodies (McAbs). The McAbs were characterized by Western blotting analysis.


RESULTS
The Plasmodium falciparum lactate dehydrogenase gene was amplified and cloned into ex pression vector pGEX-3X. The recombinant pLDH plasmid was expressed in E. coli) BL-21 cells. 15 cell lines of McAbs with high titer against pLDH were obtained using the recombinant pLDH as immunogen. Western blotting analysis showed that these McAbs recognized a Mr 33,000 of native Plasmodium falci parvum protein without cross reaction with constituents of red blood cell of febrile patients from endemic area of malaria.


CONCLUSIONS
Fifteen hybridoma cell lines secreting high titer of McAb specific to Plasmodium falciparum LDH were established based on the recombinant pLDH.
Wang
Jun-yun
JY
National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, Shanghai, 200025, China.
Bao
Yi-fang
YF
Yang
Yue-tao
YT
Tang
Lin-hua
LH
D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D000913:Antibodies, Protozoan / Q000276:immunology; D000918:Antibody Specificity / Q000276:immunology*; D001483:Base Sequence; D045744:Cell Line, Tumor; D004797:Enzyme-Linked Immunosorbent Assay; D007074:Immunoglobulin G / Q000276:immunology; D007770:L-Lactate Dehydrogenase / Q000235:genetics / Q000276:immunology*; D016778:Malaria, Falciparum / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D010963:Plasmodium falciparum / Q000201:enzymology / Q000235:genetics / Q000276:immunology; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000276:immunology / Q000302:isolation & purification
D000911:Antibodies, Monoclonal; D000913:Antibodies, Protozoan; D007074:Immunoglobulin G; D011994:Recombinant Proteins; D007770:L-Lactate Dehydrogenase