Gene gun delivered DNA-based immunizations mediate rapid production of murine monoclonal antibodies to the Flt-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 repetitive immunizations, multiple sites (RIMMS). Plasmid DNA encoding Flt-3/Fc was coated onto gold particles, which were subsequently propelled into the epidermis of mice using biolistic particle bombardment using the Accell gene gun. Pools of immune peripheral lymph node cells were somatically fused 13 days after the onset of delivery of DNA encoding the target antigen. To determine 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 presented demonstrates the successful identification and characterization of class-switched, affinity-matured MAbs that bind to the Flt-3 receptor. When compared to conventional methodologies or intramuscular targeted DNA-based immunization 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.
Kilpatrick
K E
KE
Department of Molecular Sciences, Glaxo Wellcome, Research Triangle Park, NC 27709, USA.
Cutler
T
T
Whitehorn
E
E
Drape
R J
RJ
Macklin
M D
MD
Witherspoon
S M
SM
Singer
S
S
Hutchins
J T
JT
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000917:Antibody Formation; D019470:Biolistics; D004247:DNA / Q000235:genetics / Q000276:immunology; D006801:Humans; D007114:Immunization / Q000379:methods*; D008565:Membrane Proteins / Q000235:genetics / Q000276:immunology*; D051379:Mice
D000911:Antibodies, Monoclonal; D008565:Membrane Proteins; C084881:flt3 ligand protein; D004247:DNA