4.5 Article

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

Journal

ANALYTICAL BIOCHEMISTRY
Volume 454, Issue -, Pages 38-40

Publisher

ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2014.03.011

Keywords

Buffer scouting; Regeneration; Affinity; Protein-protein interaction; High throughput; Surface plasmon resonance

Funding

  1. EFRO provinces of Gelderland and Overijssel, Netherlands

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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. (C) 2014 Elsevier Inc. All rights reserved.

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