Description
Antibodies are essential for structural determinations and functional studies of membrane proteins, but antibody generation is limited by the availability of properly-folded and purified antigen. We describe the first application of genetic immunization to a structurally diverse set of membrane proteins to show that immunization of mice with DNA alone produced antibodies against 71% (n = 17) of the bacterial and viral targets. Antibody production correlated with prior reports of target immunogenicity in host organisms, underscoring the efficiency of this DNA-gold micronanoplex approach. To generate each antigen for antibody characterization, we also developed a simple in vitro membrane protein expression and capture method. Antibody specificity was demonstrated upon identifying, for the first time, membrane-directed heterologous expression of the native sequences of the FopA and FTT1525 virulence determinants from the select agent Francisella tularensis SCHU S4. These approaches will accelerate future structural and functional investigations of therapeutically-relevant membrane proteins.
Details
Title
- Polyclonal Antibody Production for Membrane Proteins via Genetic Immunization
Contributors
- Hansen, Debra (Author)
- Robida, Mark (Author)
- Craciunescu, Felicia (Author)
- Loskutov, Andrey (Author)
- Dorner, Katerina (Author)
- Rodenberry, John-Charles (Author)
- Wang, Xiao (Author)
- Olson, Tien (Author)
- Patel, Hetal (Author)
- Fromme, Petra (Author)
- Sykes, Kathryn (Author)
- Biodesign Institute (Contributor)
- Innovations in Medicine (Contributor)
- Applied Structural Discovery (Contributor)
- College of Liberal Arts and Sciences (Contributor)
- School of Molecular Sciences (Contributor)
Date Created
The date the item was original created (prior to any relationship with the ASU Digital Repositories.)
2016-02-24
Resource Type
Collections this item is in
Identifier
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Digital object identifier: 10.1038/srep21925
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Identifier TypeInternational standard serial numberIdentifier Value2045-2322
Note
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The final version of this article, as published in Scientific Reports, can be viewed online at: https://www.nature.com/articles/srep21925
Citation and reuse
Cite this item
This is a suggested citation. Consult the appropriate style guide for specific citation guidelines.
Hansen, D. T., Robida, M. D., Craciunescu, F. M., Loskutov, A. V., Dörner, K., Rodenberry, J., . . . Sykes, K. F. (2016). Polyclonal Antibody Production for Membrane Proteins via Genetic Immunization. Scientific Reports, 6(1). doi:10.1038/srep21925