Gene gun delivered DNA-based immunizations mediate rapid production of murine monoclonal antibodies to the Fit-3 receptor

Class-switched, affinity-matured murine monoclonal antibody (MAb)-producing cell. lines were generated against the Flt-3 receptor in less than 4 weeks following polynucleotide immunizations, used in conjunction with repetitiv e immunizations, multiple sites (RIMMS), Plasmid DNA encoding Flt-3/Fc was coated onto gold particles, which were subsequently propelled into the epid ermis of mice using biolistic particle bombardment using the Accell gene gu n. Pools of immune peripheral lymph node cells were somatically fused 13 da ys after the onset of delivery of DNA encoding the target antigen. To deter mine if early responses could be augmented, DNA-encoding murine GM-CSF was delivered 3 days prior to the Flt-3/Fc DNA immunizations. The data presente d demonstrates the successful identification and characterization of class- switched, affinity-matured MAbs that bind to the Flt-3 receptor. When compa red to conventional methodologies or intramuscular targeted DNA-based immun ization for the generation of MAbs, use of the gene gun in conjunction with RIMMS allows for a more rapid production of affinity-matured MAb-producing cell lines.