[Preparation of monoclonal antibodies against malaria through immunization of mice with recombinant plasmid DNA].
Xie W, Ping Zhong, Pan W, Q Chen, Q Wang, Danfeng Lu
Abstract
Xie W, Ping Zhong, Pan W, Q Chen, Q Wang, Danfeng Lu
Abstract
BALB/c mice were immunized by injection of recombinant plasmid DNA carrying MSP1-31 fragment of Plasmodium falciparum. After antiserum was induced, McAbs were prepared by fusing SP2/0 myeloma cells with spleen cells. Specific McAbs were detected in hybridoma culture supernatant and mouse ascites by ELISA, positive hybridomas were cloned by technique of limiting dilutions. Two McAbs, 9H9 and 8A2, against the malaria protein were produced and characterized. The titers of the two McAbs in acites were 1:10,000 and 1:2500 respectively. The Ig subclass of these McAbs was IgG1. Western blot test showed that the McAb specifically reacted with MSP1-31 antigen. These results proved that monoclonal antibodies could be produced by use of immunization of mice with plasmid DNA or DNA vaccine.
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BALB/c mice were immunized by injection of recombinant plasmid DNA carrying MSP1-31 fragment of Plasmodium falciparum. After antiserum was induced, McAbs were prepared by fusing SP2/0 myeloma cells with spleen cells. Specific McAbs were detected in hybridoma culture supernatant and mouse ascites by ELISA, positive hybridomas were cloned by technique of limiting dilutions. Two McAbs, 9H9 and 8A2, against the malaria protein were produced and characterized. The titers of the two McAbs in acites were 1:10,000 and 1:2500 respectively. The Ig subclass of these McAbs was IgG1. Western blot test showed that the McAb specifically reacted with MSP1-31 antigen. These results proved that monoclonal antibodies could be produced by use of immunization of mice with plasmid DNA or DNA vaccine.
Key concepts: Recombinant DNA, Monoclonal antibody, Virology, Molecular biology, Titer, Biology, Plasmid, Antigen