Generation of High-Specificity Antibodies against Membrane Proteins Using DNA-Gold Micronanoplexes for Gene Gun Immunization

Debra Hansen, Felicia M. Craciunescu, Petra Fromme, Stephen Johnston, Kathryn F. Sykes

Research output: Contribution to journalArticle

2 Scopus citations

Abstract

Membrane proteins are the molecular interface of the cell and its environs; however, studies of membrane proteins are highly technically challenging, mainly due to instability of the isolated protein. Towards the production of antibodies that recognize properly folded and stabilized forms of membrane protein antigen, we describe a DNA-based immunization method for mice that expresses the antigen in the membranes of dendritic cells, thus allowing direct presentation to the immune system. This genetic immunization approach employs a highly efficient method of biolistic delivery based on DNA-gold micronanoplexes, which are complexes of micron-sized gold particles that allow dermal penetration and nanometer-sized gold particles that provide a higher surface area for DNA binding than micron gold alone. In contrast to antibodies derived from immunizations with detergent-solubilized protein or with protein fragments, antibodies from genetic immunization are expected to have a high capacity for binding conformational epitopes and for modulating membrane protein activity.

Original languageEnglish (US)
Pages (from-to)29.20.1-29.20.22
JournalCurrent Protocols in Protein Science
Volume91
DOIs
StatePublished - Feb 21 2018

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Keywords

  • antibodies
  • conformational epitopes
  • DNA immunization
  • gene gun
  • genetic immunization
  • membrane protein

ASJC Scopus subject areas

  • Structural Biology
  • Biochemistry

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