Fluorescent aptasensor based on D-AMA/F-CSC for the sensitive and specific recognition of myoglobin

Spectrochim Acta A Mol Biomol Spectrosc. 2020 Mar 5:228:117714. doi: 10.1016/j.saa.2019.117714. Epub 2019 Nov 1.

Abstract

A novel fluorescent biosensor based on dabcyl [(E)-4-((4-(dimethylamino) phenyl) diazenyl)benzoic acid] -modified anti-Mb aptamer (D-AMA) and 6-FAM(6-carboxyfluorescein) -modified complementary short chain (F-CSC)for the specific and sensitive detection of Mb levels is presented in this study. In PBS buffer solution, D-AMA bound to F-CSC, and then dabcyl quenched the fluorescence of 6-FAM. After adding Mb into the system, D-AMA bound to Mb and separated from F-CSC. The fluorescence of 6-FAM was restored after it separated from dabcyl. The assay exhibited high specificity and sensitivity toward Mb, with a low limit of detection of 0.07 ng/mL (S/N = 3) and linear relationships of 0.1-5 ng/mL. It was further applied to detect Mb levels in spiked human blood sera samples.

Keywords: 6-Carboxyfluorescein; Aptasensor; Dabcyl; Fluorescence; Myoglobin.

MeSH terms

  • Animals
  • Aptamers, Nucleotide / chemistry*
  • Base Sequence
  • Biosensing Techniques*
  • Cattle
  • DNA / analysis
  • Fluoresceins / chemistry*
  • Humans
  • Myoglobin / blood*
  • Spectrometry, Fluorescence

Substances

  • Aptamers, Nucleotide
  • Fluoresceins
  • Myoglobin
  • 6-carboxyfluorescein
  • DNA