High-throughput and multiplexed regeneration buffer scouting for affinity-based interactions

Anal Biochem. 2014 Jun 1;454:38-40. doi: 10.1016/j.ab.2014.03.011. Epub 2014 Mar 21.

Abstract

Affinity-based analyses on biosensors depend partly on regeneration between measurements. Regeneration is performed with a buffer that efficiently breaks all interactions between ligand and analyte while maintaining the active binding site of the ligand. We demonstrated a regeneration buffer scouting using the combination of a continuous flow microspotter with a surface plasmon resonance imaging platform to simultaneously test 48 different regeneration buffers on a single biosensor. Optimal regeneration conditions are found within hours and consume little amounts of buffers, analyte, and ligand. This workflow can be applied to any ligand that is coupled through amine, thiol, or streptavidin immobilization.

Keywords: Affinity; Buffer scouting; High throughput; Protein–protein interaction; Regeneration; Surface plasmon resonance.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amines / chemistry
  • Buffers
  • Ligands
  • Streptavidin / chemistry
  • Sulfhydryl Compounds / chemistry
  • Surface Plasmon Resonance / methods*
  • Time Factors

Substances

  • Amines
  • Buffers
  • Ligands
  • Sulfhydryl Compounds
  • Streptavidin