High efficiency acetylcholinesterase immobilization on DNA aptamer modified surfaces

Molecules. 2014 Apr 21;19(4):4986-96. doi: 10.3390/molecules19044986.

Abstract

We report here the in vitro selection of DNA aptamers for electric eel acetylcholinesterase (AChE). One selected aptamer sequence (R15/19) has a high affinity towards the enzyme (Kd=157±42 pM). Characterization of the aptamer showed its binding is not affected by low ionic strength (~20 mM), however significant reduction in affinity occurred at high ionic strength (~1.2 M). In addition, this aptamer does not inhibit the catalytic activity of AChE that we exploit through immobilization of the DNA on a streptavidin-coated surface. Subsequent immobilization of AChE by the aptamer results in a 4-fold higher catalytic activity when compared to adsorption directly on to plastic.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylcholinesterase / chemistry*
  • Animals
  • Aptamers, Nucleotide / chemistry*
  • Base Sequence
  • Electrophorus
  • Enzymes, Immobilized / chemistry*
  • Fish Proteins / chemistry*
  • Kinetics
  • Molecular Sequence Data
  • Nucleic Acid Conformation
  • Osmolar Concentration
  • Protein Binding
  • SELEX Aptamer Technique
  • Streptavidin / chemistry

Substances

  • Aptamers, Nucleotide
  • Enzymes, Immobilized
  • Fish Proteins
  • Streptavidin
  • Acetylcholinesterase