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

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

Author information

1
Downstream Processing, Synthon Biopharmaceuticals, 6503 GN Nijmegen, The Netherlands; Bioprocess Engineering Group, Wageningen University, 6700 EV Wageningen, The Netherlands. Electronic address: karin.geuijen@synthon.com.
2
Medical Cell Biophysics Group, MIRA Institute, Faculty of Science and Technology, University of Twente, 7500 AE Enschede, The Netherlands; IBIS Technologies, 7521 PR Enschede, The Netherlands.
3
Bioprocess Engineering Group, Wageningen University, 6700 EV Wageningen, The Netherlands.
4
Downstream Processing, Synthon Biopharmaceuticals, 6503 GN Nijmegen, The Netherlands; Bioprocess Engineering Group, Wageningen University, 6700 EV Wageningen, The Netherlands.

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

PMID:
24657361
DOI:
10.1016/j.ab.2014.03.011
[Indexed for MEDLINE]

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