PMID: 16526410Mar 11, 2006Paper

Genetic immunization for antibody generation in research animals by intravenous delivery of plasmid DNA

BioTechniques
Mary K BatesHans Herweijer

Abstract

Genetic immunization is an attractive approach to generate antibodies because native proteins are expressed in vivo with normal posttranscriptional modifications, avoiding time-consuming and costly antigen isolation or synthesis. Hydrodynamic tail or limb vein delivery of naked plasmid DNA expression vectors was used to induce antigen-specific antibodies in mice, rats, and rabbits. Both methods allowed the efficient generation of high-titer, antigen-specific antibodies with an overall success rate of Western detectable antibodies of 78% and 92%, respectively. High-titer antibodies were typically present after 3 hydrodynamic tail vein plasmid DNA deliveries, 5 weeks after the initial injection (i.e., prime). For hydrodynamic limb vein plasmid DNA delivery, two deliveries were sufficient to induce high-titer antibody levels. Tail vein delivery was less successful at generating antibodies directed against secreted proteins as compared with limb vein delivery. Material for screening was generated by,transfection of the immunization vector into mammalian cell lines. The cell line (COS-7) that produced the highest level of antigen expression performed best in Western blot analysis screens. In summary, intravenous delivery of antigen-...Continue Reading

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Citations

Feb 9, 2011·Journal of Molecular Medicine : Official Organ of the Gesellschaft Deutscher Naturforscher Und Ärzte·Orestis ArgyrosRichard P Harbottle
Jan 20, 2007·Molecular Therapy : the Journal of the American Society of Gene Therapy·Zane C NealHans Herweijer
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Nov 5, 2013·Bioanalysis·Denise M O'Hara, Valerie Theobald
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Dec 15, 2006·Gene Therapy·H Herweijer, J A Wolff
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Jan 1, 2011·Current Protocols in Mouse Biology·Christine I WooddellGuofeng Zhang
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May 16, 2021·Protein Engineering, Design & Selection : PEDS·Kasandra Bélanger, Jamshid Tanha
Jun 6, 2009·Cell Metabolism·Jason T DeanJames C Liao

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