Ratiometric Fluorescence Aptasensor of Allergen Protein Based on Multivalent Aptamer-Encoded DNA Flowers as Fluorescence Resonance Energy Transfer Platform

Anal Chem. 2024 May 7;96(18):6947-6957. doi: 10.1021/acs.analchem.3c05894. Epub 2024 Apr 24.

Abstract

Life-threatening allergic reactions to food allergens, particularly peanut protein Ara h1, are a growing public health concern affecting millions of people worldwide. Thus, accurate and rapid detection is necessary for allergen labeling and dietary guidance and ultimately preventing allergic incidents. Herein, we present a novel ratiometric fluorescence aptasensor based on multivalent aptamer-encoded DNA flowers (Mul-DNFs) for the high-stability and sensitive detection of allergen Ara h1. The flower-shaped Mul-DNFs were spontaneously packaged using ultralong polymeric DNA amplicons driven by a rolling circle amplification reaction, which contains a large number of Ara h1 specific recognition units and has excellent binding properties. Furthermore, dual-color fluorescence-labeled Mul-DNFs probes were developed by hybridizing them with Cy3- and Cy5-labeled complementary DNA (cDNA) to serve as a ratiometric fluorescence aptasensor platform based on fluorescence resonance energy transfer. Benefiting from the combined merits of the extraordinary synergistic multivalent binding ability of Mul-DNFs, the excellent specificity of the aptamer, and the sensitivity of the ratiometric sensor to avoid exogenous interference. The developed ratiometric aptasensor showed excellent linearity (0.05-2000 ng mL-1) with a limit of detection of 0.02 ng mL-1. Additionally, the developed ratiometric fluorescence aptasensor was utilized for quantifying the presence of Ara h1 in milk, infant milk powder, cookies, bread, and chocolate with recoveries of 95.7-106.3%. The proposed ratiometric aptasensor is expected to be a prospective universal aptasensor platform for the rapid, sensitive, and accurate determination of food and environmental hazards.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Allergens* / analysis
  • Animals
  • Antigens, Plant* / analysis
  • Aptamers, Nucleotide* / chemistry
  • Biosensing Techniques / methods
  • DNA / chemistry
  • Fluorescence Resonance Energy Transfer*
  • Fluorescent Dyes / chemistry
  • Glycoproteins / analysis
  • Glycoproteins / chemistry
  • Limit of Detection
  • Membrane Proteins*
  • Plant Proteins / analysis
  • Plant Proteins / chemistry

Substances

  • Aptamers, Nucleotide
  • Allergens
  • Antigens, Plant
  • Ara h 1 protein, Arachis hypogaea
  • DNA
  • Glycoproteins
  • Fluorescent Dyes
  • Plant Proteins
  • Membrane Proteins