Development of a synthetic receptor protein for sensing inflammatory mediators interferon‐γ and tumor necrosis factor‐α
Abstract
Intestinal inflammation has been implicated in a number of diseases, including diabetes, Crohn's disease, and irritable bowel syndrome. Important components of inflammation are interferon‐γ (IFN‐γ) and tumor necrosis factor‐α (TNF‐α), which are elevated both on the luminal and submucosal sides of the intestinal epithelial barrier in several diseases. Here, we developed a novel Escherichia coli based detection system for IFN‐γ and TNF‐α comprised of a chimeric protein and a simple signal transduction construct, which could be deployed on the luminal side of the intestine. OmpA of E. coli was engineered to detect IFN‐γ or TNF‐α through the replacement of extracellular loops with peptide fragments from OprF of P. aeruginosa. OmpA/OprF chimeras were developed, capable of binding IFN‐γ or TNF‐α. The specific peptide fragments that bind IFN‐γ were identified. IFN‐γ or TNF‐α binding the OmpA/OprF chimera induced the pspA promoter, driving β‐galactosidase production. The OmpA/OprF chimera had a detection limit of 300 pM for IFN‐γ and 150 pM for TNF‐α. This work will further the development of bacteria based therapeutics for the treatment of inflammatory diseases of the gut. Biotechnol. Bioeng. 2016;113: 492–500. © 2015 Wiley Periodicals, Inc.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Jan 15, 2016
- Source ID
- 10.1002/bit.25832
Entities
People
- John C. March
- T. Christopher Aurand
Organizations
- Cornell University
- Defense Threat Reduction Agency
- National Institutes of Health