A label-free PFP-based photoelectrochemical biosensor for highly sensitive detection of PARP-1 activity

Biosens Bioelectron. 2019 Aug 1:138:111308. doi: 10.1016/j.bios.2019.05.013. Epub 2019 May 9.

Abstract

Poly(ADP-ribose) polymerase-1 (PARP-1), as an original tumor marker, has aroused wide attention in recent years. However, only a few researches have been done for PARP-1 activity detection because PARP-1 is lack of optical or electrochemical property. In this work, a label-free and high-sensitive photoelectrochemical (PEC) biosensor for PARP-1 activity detection based on poly[9,9-bis(6'-N,N,N-trimethylammonium)hexyl]fluorenylene phenylene (PFP) has been designed. To the best of our knowledge, it is the first time that PEC has been used to monitor PARP-1 activity. PARP-1 were activated under the function of activated dsDNA, as a result, branched polymers of ADP-ribose (PAR) with plentiful negative charge were formed in the presence of nicotinamide adenine dinucleotide (NAD+). Subsequently, positively charged PFP with good photoelectrochemical properties, were absorbed on PAR via electrostatic interaction. High photocurrent was produced under light induction, which was depended on the PARP-1 activity. The biosensor has a wide linear range from 0.01 to 2 U with a detection limit of 0.007 U. The strategy has been applied in breast and ovarian cancer cells to detection PARP-1 activity with approving results, which signifies that it is a promising tool for clinical diagnosis.

Keywords: Biosensor; Detection; Electrostatic interaction; Enzyme activity; Poly(ADP-Ribose) polymerase-1; Poly[9,9-bis(6′-N,N,N-trimethylammonium)hexyl]fluorenylene phenylene.

MeSH terms

  • Biomarkers, Tumor / metabolism*
  • Biosensing Techniques*
  • Bis-Trimethylammonium Compounds / chemistry*
  • Cell Line, Tumor
  • Electrochemical Techniques
  • Electrodes
  • Enzyme Assays / methods
  • Humans
  • Indoles / pharmacology
  • Limit of Detection
  • NAD / pharmacology
  • Photochemical Processes
  • Poly (ADP-Ribose) Polymerase-1 / metabolism*
  • Poly(ADP-ribose) Polymerase Inhibitors / pharmacology
  • Static Electricity

Substances

  • Biomarkers, Tumor
  • Bis-Trimethylammonium Compounds
  • Indoles
  • Poly(ADP-ribose) Polymerase Inhibitors
  • poly(9,9-bis(6'-N,N,N-trimethylammonium)hexyl)fluorenylene phenylene
  • NAD
  • rucaparib
  • Poly (ADP-Ribose) Polymerase-1