Gene Gun Delivered DNA-Based Immunizations Mediate Rapid Production of Murine Monoclonal Antibodies to the Flt-3 Receptor
- 1 December 1998
- journal article
- research article
- Published by Mary Ann Liebert Inc in Hybridoma
- Vol. 17 (6), 569-576
- https://doi.org/10.1089/hyb.1998.17.569
Abstract
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.Keywords
This publication has 23 references indexed in Scilit:
- Rapid Development of Affinity Matured Monoclonal Antibodies Using RIMMSHybridoma, 1997
- A Generic Method for the Production of Cell Lines Expressing High Levels of 7-Transmembrane ReceptorsAnalytical Biochemistry, 1997
- Generation of a monoclonal antibody to a defined portion of the Helicobacter pylori vacuolating cytotoxin by DNA immunizationJournal of Biotechnology, 1996
- DNA–based immunization by in vivo transfection of dendritic cellsNature Medicine, 1996
- DNA vaccines: A novel approach to immunizationInternational Journal of Immunopharmacology, 1995
- Examination of Parameters Affecting the Elicitation of Humoral Immune Responses by Particle Bombardment-Mediated Genetic ImmunizationDNA and Cell Biology, 1993
- Genetic immunization is a simple method for eliciting an immune responseNature, 1992
- In vivo and in vitro gene transfer to mammalian somatic cells by particle bombardment.Proceedings of the National Academy of Sciences of the United States of America, 1990
- Langerhans' cells, veiled cells, and interdigitating cells in the mouse recognized by a monoclonal antibody.The Journal of Experimental Medicine, 1986
- A cell surface ELISA in the mouse using only poly-l-lysine as cell fixativeJournal of Immunological Methods, 1985