The characterization and validation of 17β-estradiol binding aptamers

J Steroid Biochem Mol Biol. 2017 Mar:167:14-22. doi: 10.1016/j.jsbmb.2016.09.018. Epub 2016 Sep 23.

Abstract

The rapid and sensitive detection of small molecules is garnering increasing importance, and aptamers show great promise in replacing expensive, elaborate detection platforms exploiting chromatographic separation or antibody-based assays. The characterization of aptamer interaction with small molecule targets is not facile, and there is a mature need for a rapid, high-throughput technique for the analysis of aptamer-small molecule kinetics and affinity. In this work we present methodologies for the evaluation of aptamer-small molecule interactions, using the aptamers reported against the steroid 17β-estradiol as a model system. Microscale thermophoresis, apta-PCR affinity assay and surface plasmon resonance were explored to evaluate the reported aptamers' binding properties in terms of affinity and specificity, and were demonstrated to be successfully applied to the analysis of aptamer-small molecule interactions.

Keywords: 17β-estradiol; Apta-PCR affinity assay (APAA); Aptamers; Microscale thermophoresis (MST); Small molecule; Surface Plasmon Resonance (SPR).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aptamers, Nucleotide / metabolism*
  • Binding Sites
  • Dose-Response Relationship, Drug
  • Estradiol / metabolism*
  • G-Quadruplexes
  • Humans
  • Interferometry
  • Kinetics
  • Ligands
  • Magnetics
  • Nucleic Acid Conformation
  • Polymerase Chain Reaction
  • Progesterone / chemistry
  • Protein Binding
  • SELEX Aptamer Technique / methods*
  • Sensitivity and Specificity
  • Surface Plasmon Resonance

Substances

  • Aptamers, Nucleotide
  • Ligands
  • Progesterone
  • Estradiol