Generation of high titer antisera in rabbits by DNA immunization.
Mesothelin is a GPI-linked, membrane-associated differentiation antigen that is over-expressed in several forms of human cancers. Intradermal injection into rabbits of plasmid DNA encoding full length mesothelin resulted in antisera with titers as high as 1:100,000. Each immunization consisted of 320 microg of DNA delivered into 4 sites. After the initial three injections antisera titers were moderate (between 10 to 30,000) and fell over the course of about 7 weeks. When the titers had fallen, an injection of a booster dose of DNA resulted in very high titers of antisera. These antisera contained IgGs that could bind to both recombinant mesothelin made in Eschericha coli and to mesothelin present on human cells in Western blots and in immunofluorescence assays. These observations indicate that simple intradermal DNA immunization of rabbits can result in high titers of antibodies that can be used for a variety of purposes.
Laboratory of Molecular Biology, Division of Basic Sciences, National Cancer Institute, National Institutes of Health, Building 37, Room 4E16, 37 Convent Dr Msc 4255, Bethesda, MD 20892-4255, USA.
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PS
Chowdhury
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Gallo
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Pastan
D000818:Animals; D000906:Antibodies / Q000276:immunology* / Q000302:isolation & purification*; D004247:DNA / Q000276:immunology*; D006801:Humans; D007114:Immunization / Q000379:methods; D011817:Rabbits
D000906:Antibodies; D004247:DNA