Selection and Application of Aptamer Affinity for Protein Purification

Methods Mol Biol. 2022:2466:187-203. doi: 10.1007/978-1-0716-2176-9_13.

Abstract

Aptamers are affinity-based oligonucleotide ligands raised against a target molecule, which might be of proteic or other nature. Aptamers are developed by using a reiterative in vitro selection procedure, named SELEX, in which the target is exposed to a combinatorial oligonucleotide combinatorial library. Target bound oligonucleotides are eluted, and PCR amplified followed by the next SELEX round. The process is repeated until no further increase in target binding affinity and specificity is achieved. Selected aptamers are identified and immobilized for protein purification. In view of their stability against denaturation and capability of renaturation, low costs of production, easiness of modification and stabilization, oligonucleotide aptamers are excellent tools as high-affinity ligands for applications of protein purification.

Keywords: Aptamer affinity; Aptamer–protein complexes; Electrophoretic mobility shift assay; Immobilized aptamer; Protein purification; Pull down-assay.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aptamers, Nucleotide* / chemistry
  • Gene Library
  • Ligands
  • Polymerase Chain Reaction
  • SELEX Aptamer Technique* / methods

Substances

  • Aptamers, Nucleotide
  • Ligands