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Author Notes:

Corresponding author. *E‐mail: d.overby@imperial.ac.uk, dan.stamer@duke.edu

The authors thank Dr. S. Ladame, Dr. E.T. Pashuck, Dr. Y. Lin, and Dr. R. Chandrawatri for helpful discussions regarding free radical chemistry, peptide synthesis, and fluorescent particle functionalization. K. M. Perkumas and N. E.

Ashpole for their assistance with the immunostaining of cells and setting up the Ibidi shear system.

Dr. C. Silvestri and Dr. E. Reina‐Torres for proof reading the manuscript.

J.Y.H.C. acknowledges the PhD Studentship from Imperial College London Department of Bioengineering.

JYHC, DRO, LWC and WDS are named inventors on a pending patent covering the technology described within the manuscript.

Subjects:

Research Funding:

D.R.O. and W.D.S. gratefully acknowledge the funding support from NIH Grants (EY022359, EY005722) and Research to Prevent Blindness Foundation.

W.M.D. thanks the funding support from NIH Grant (F32 EY023468) and Research to Prevent Blindness Foundation.

M.M.S. and L.W.C. thank the Medical Engineering Solutions in the Osteoarthritis Centre of Excellence, funded by the Wellcome Trust and Engineering and Physical Sciences Research Council (EPSRC; 088844).

M.M.S. would also like to acknowledge the funding support from the ERC Seventh Framework Programme Consolidator grant “Naturale CG” under grant agreement no. 616417.

Keywords:

  • Science & Technology
  • Technology
  • Engineering, Biomedical
  • Nanoscience & Nanotechnology
  • Materials Science, Biomaterials
  • Engineering
  • Science & Technology - Other Topics
  • Materials Science
  • endothelial cells
  • immunoassays
  • nitric oxide detection
  • peptide biosensors
  • peroxynitrite
  • PROTEIN-TYROSINE NITRATION
  • TUMOR MICROVASCULAR PERMEABILITY
  • SOLUBLE GUANYLATE-CYCLASE
  • SHEAR-STRESS
  • MASS-SPECTROMETRY
  • OXIDATIVE STRESS
  • PROSTACYCLIN SYNTHASE
  • BIOLOGICAL-SYSTEMS
  • SUPEROXIDE
  • RELEASE

Peptide-Functionalized Fluorescent Particles for In Situ Detection of Nitric Oxide via Peroxynitrite-Mediated Nitration

Tools:

Journal Title:

Advanced Healthcare Materials

Volume:

Volume 6, Number 16

Publisher:

, Pages 1700383-1700383

Type of Work:

Article | Final Publisher PDF

Abstract:

Nitric oxide (NO) is a free radical signaling molecule that plays a crucial role in modulating physiological homeostasis across multiple biological systems. NO dysregulation is linked to the pathogenesis of multiple diseases; therefore, its quantification is important for understanding pathophysiological processes. The detection of NO is challenging, typically limited by its reactive nature and short half-life. Additionally, the presence of interfering analytes and accessibility to biological fluids in the native tissues make the measurement technically challenging and often unreliable. Here, a bio-inspired peptide-based NO sensor is developed, which detects NO-derived oxidants, predominately peroxynitrite-mediated nitration of tyrosine residues. It is demonstrated that these peptide-based NO sensors can detect peroxynitrite-mediated nitration in response to physiological shear stress by endothelial cells in vitro. Using the peptide-conjugated fluorescent particle immunoassay, peroxynitrite-mediated nitration activity with a detection limit of ≈100 × 10−9m is detected. This study envisions that the NO detection platform can be applied to a multitude of applications including monitoring of NO activity in healthy and diseased tissues, localized detection of NO production of specific cells, and cell-based/therapeutic screening of peroxynitrite levels to monitor pronitroxidative stress in biological samples.

Copyright information:

© 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

This is an Open Access work distributed under the terms of the Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0/).
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