2',4'-BNA/LNA aptamers: CE-SELEX using a DNA-based library of full-length 2'-O,4'-C-methylene-bridged/linked bicyclic ribonucleotides

Bioorg Med Chem Lett. 2013 Mar 1;23(5):1288-92. doi: 10.1016/j.bmcl.2012.12.093. Epub 2013 Jan 16.

Abstract

DNA-based aptamers that contain 2'-O,4'-C-methylene-bridged/linked bicyclic ribonucleotides (B/L nucleotides) over the entire length were successfully obtained using a capillary electrophoresis systematic evolution of ligands by exponential enrichment (CE-SELEX) method. A modified DNA library was prepared with an enzyme mix of KOD Dash and KOD mutant DNA polymerases. Forty 2'-O,4'-C-methylene bridged/locked nucleic acid (2',4'-BNA/LNA) aptamers were isolated from an enriched pool and classified into six groups according to their sequence. 2',4'-BNA/LNA aptamers of groups V and VI bound human thrombin with K(d) values in the range of several 10 nanomolar levels.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aptamers, Nucleotide / chemistry*
  • Aptamers, Nucleotide / genetics
  • Aptamers, Nucleotide / metabolism
  • Bridged-Ring Compounds / chemistry*
  • DNA-Directed DNA Polymerase / chemistry
  • DNA-Directed DNA Polymerase / genetics
  • Gene Library
  • Humans
  • Nucleotides / chemistry*
  • Oligonucleotides / chemistry*
  • Ribonucleotides / chemistry*
  • Ribonucleotides / genetics
  • SELEX Aptamer Technique / methods*

Substances

  • 2',4'-BNA(NC)
  • Aptamers, Nucleotide
  • Bridged-Ring Compounds
  • Nucleotides
  • Oligonucleotides
  • Ribonucleotides
  • locked nucleic acid
  • DNA-Directed DNA Polymerase