Transfection Optimization for Expression of the Recombinant Monoclonal Antibody against Plasmodium falciparum Lactate Dehydrogenase in Mammalian Cells
Keywords:
Recombinant Plasmodium falciparun monoclonal antibody, 293F cells, transfection, Plasmodium falciparum lactate dehydrogenaseAbstract
Recombinant plasmid pcDNA-pLDHAbHCγLCκ/C10 containing immunoglobulin gene encoding monoclonal antibody against lactate dehydrogenase enzyme of Plasmodium falciparum (pLDH) has been developed in our previous study. This study was aimed to optimize the condition of transfection reagent for experssion of thee crombinant monoclonal antibody against pLDH in 293F cs.e ll By optimizing transfection condition for a commercial pEGFP-N1 plasmid to transfection reagent, the optimal ratiowas at 1 μg:1.2 μl in 293F cells. By this ratio, the plasmid pcDNA-pLDHAbHCγLCκ/C10 was also transfected into 293F cells to express the recombinant monoclonal antibody and maximal level was found in culture supernatant at day 4 as determined by ELISA. The recombinant monoclonal antibody protein pattern was similar to that of the mouse monoclonal antibody against pLDH as analyzed by SDS-PAGE and Western blotting. Furthermore, this monoclonal antibody could react specifically to pLDH antigen in P. falciparum infected cells by IFA. Thus, the recombinant monoclonal antibody had potential in development for P. faciparum malarial diagnosis
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