Preparation of Monoclonal Antibody Based on Single Intrasplenic Immunization of Plasmid DNA
质粒DNA单次脾内注射制备单克隆抗体

To set up a new rapid and efficient way for the preparation of specific monoclonal antibodies (MAbs) with plasmid DNA immunization. METHODS: The fusion gene of IL-2 signal peptide and profilin 1 by 'overlapping PCR' was obtained and inserted into the vector pVAX1 to construct the recombinant plasmid pVAX-IL2-profl. Another fusion gene of IgG kappa chain signal peptide and profilin 1 by 'no template PCR' was obtained and inserted into the vector pVAX1 to construct the recombinant plasmid pVAX-Igkappa-prof1. BALB/c mice were single intrasplenic immunized with plasmid DNA. Results After cell fusion and hybridomas screening by indirect ELISA, we charactered two mAbs (1D8A2B4 and 4B12A5E3) against profilin 1. The MAbs isotype were determined as IgM and IgG3. CONCLUSION: A single intrasplenic plasmid DNA immunization is rapid and efficient and can be used as a helpful tool for the production of specific mAb.