High-affinity monoclonal antibodies to PED/PEA-15 generated using 5 microg of DNA.
Class-switched, affinity-matured murine monoclonal antibody (MAb) producing cell lines reactive with PED/PEA-15 were generated and isolated in less than 4 weeks following polynucleotide immunizations using only 5 microg of DNA in conjunction with the Powderject gene gun. Somatic fusions of peripheral lymph node cells were performed 13 days after initiating delivery of DNA encoding the target antigen. The data presented demonstrates the rapid production, identification, and characterization of class-switched, affinity-matured MAbs that bind PED/PEA-15. The reported strategy enabled the rapid development of MAbs that are useful in enzyme-linked immunoadsorbent assay (ELISA), Western blotting, and immunoprecipitations.
Kilpatrick
K E
KE
Department of Protein Sciences, Glaxo Welcome, Inc., Research Triangle Park, NC 27709,USA. kek17850@glaxowellcome.com
Danger
D P
DP
Hull-Ryde
E A
EA
Dallas
W
W
D000818:Animals; D000974:Antibodies, Antinuclear / Q000096:biosynthesis* / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D000915:Antibody Affinity / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology; D051017:Apoptosis Regulatory Proteins; D015153:Blotting, Western; D004247:DNA / Q000276:immunology*; D017931:DNA Primers / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D015395:Histocompatibility Antigens Class I / Q000235:genetics* / Q000276:immunology; D006801:Humans; D007114:Immunization / Q000379:methods; D047908:Intracellular Signaling Peptides and Proteins; D051379:Mice; D010750:Phosphoproteins / Q000235:genetics* / Q000276:immunology; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins
D000974:Antibodies, Antinuclear; D000911:Antibodies, Monoclonal; D051017:Apoptosis Regulatory Proteins; D017931:DNA Primers; D015395:Histocompatibility Antigens Class I; D047908:Intracellular Signaling Peptides and Proteins; C092039:PEA15 protein, human; C490444:Pea15 protein, mouse; D010750:Phosphoproteins; C032907:Q surface antigens; D011993:Recombinant Fusion Proteins; D004247:DNA