Affinity purification of bacterial outer membrane vesicles (OMVs) utilizing a His-tag mutant

Nathan Alves, Kendrick B. Turner, Kyle A. DiVito, Michael A. Daniele, Scott A. Walper

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

To facilitate the rapid purification of bacterial outer membrane vesicles (OMVs), we developed two plasmid constructs that utilize a truncated, transmembrane protein to present an exterior histidine repeat sequence. We chose OmpA, a highly abundant porin protein, as the protein scaffold and utilized the lac promoter to allow for inducible control of the epitope-presenting construct. OMVs containing mutant OmpA-His6 were purified directly from Escherichia coli culture media on an immobilized metal affinity chromatography (IMAC) Ni-NTA resin. This enabling technology can be combined with other molecular tools directed at OMV packaging to facilitate the separation of modified/cargo-loaded OMV from their wt counterparts. In addition to numerous applications in the pharmaceutical and environmental remediation industries, this technology can be utilized to enhance basic research capabilities in the area of elucidating endogenous OMV function.

Original languageEnglish (US)
Pages (from-to)139-146
Number of pages8
JournalResearch in Microbiology
Volume168
Issue number2
DOIs
StatePublished - Feb 1 2017

Fingerprint

Membranes
Technology
Porins
Proteins
Product Packaging
Affinity Chromatography
Histidine
Culture Media
Epitopes
Industry
Plasmids
Metals
Escherichia coli
Research
Pharmaceutical Preparations

Keywords

  • Affinity purification
  • Escherichia coli
  • Extracellular vesicles
  • His-tag
  • IMAC
  • Outer membrane vesicles (OMVs)

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

Affinity purification of bacterial outer membrane vesicles (OMVs) utilizing a His-tag mutant. / Alves, Nathan; Turner, Kendrick B.; DiVito, Kyle A.; Daniele, Michael A.; Walper, Scott A.

In: Research in Microbiology, Vol. 168, No. 2, 01.02.2017, p. 139-146.

Research output: Contribution to journalArticle

Alves, Nathan ; Turner, Kendrick B. ; DiVito, Kyle A. ; Daniele, Michael A. ; Walper, Scott A. / Affinity purification of bacterial outer membrane vesicles (OMVs) utilizing a His-tag mutant. In: Research in Microbiology. 2017 ; Vol. 168, No. 2. pp. 139-146.
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