Selection of DNA aptamers against penicillin G using Capture-SELEX for the development of an impedimetric sensor

Talanta. 2017 Jan 1:162:232-240. doi: 10.1016/j.talanta.2016.09.058. Epub 2016 Sep 28.

Abstract

This paper describes for the first time the selection of aptamers selective to penicillin. Aptamers were selected using a specific process called Capture-SELEX (Systematic Evolution of Ligands by Exponential Enrichment). This technique is based on the selection of DNA aptamers using penicillin G in solution while the ssDNA library is fixed on a support. One aptamer showing a good affinity to penicillin was finally selected and tested in electrochemical sensor configuration, using electrochemical impedance spectroscopy as detection technique. The developed aptasensor allowed the detection of penicillin in a wide concentration range, comprised between 0.4 and 1000µgL-1 Such performance was compatible with milk analysis, as the maximum residue limit tolerated in this matrix is 4µgL-1. The selectivity of the developed sensor was also studied, showing that the sensor was also able to bind other beta-lactam antibiotics, although with a weaker affinity. Finally the sensor was used for detection of penicillin G in milk. It was shown that a simple sample treatment with isopropanol followed by filtration was sufficient to eliminate matrix effects, allowing the determination of penicillin in milk at concentrations compatible with legislation requirements.

Keywords: Aptamers; Capture-SELEX; Impedimetric sensor; Milk analysis; Penicillin.

MeSH terms

  • Animals
  • Aptamers, Nucleotide / chemistry*
  • Aptamers, Nucleotide / genetics
  • Aptamers, Nucleotide / metabolism
  • Base Sequence
  • Binding, Competitive
  • Biosensing Techniques / methods*
  • DNA, Single-Stranded / chemistry
  • DNA, Single-Stranded / genetics
  • DNA, Single-Stranded / metabolism
  • Dielectric Spectroscopy / methods*
  • Milk / chemistry
  • Penicillin G / analysis
  • Penicillin G / chemistry*
  • Penicillin G / metabolism
  • Reproducibility of Results
  • SELEX Aptamer Technique / methods*
  • beta-Lactams / analysis
  • beta-Lactams / chemistry
  • beta-Lactams / metabolism

Substances

  • Aptamers, Nucleotide
  • DNA, Single-Stranded
  • beta-Lactams
  • Penicillin G