Comparison of fluorescence labelling techniques for the selection of affinity ligands from solid-phase combinatorial libraries

Ana S. Pina, Christopher R. Lowe, Ana Cecília A. Roque

Research output: Contribution to journalConference articlepeer-review

2 Citations (Scopus)

Abstract

This study reports the comparison of fluorimetric techniques (fluorescence microscopy and spectrofluorimetry on a 96-well format) for the on-bead screening of combinatorial libraries of affinity ligands for chromatographic separations. Two solid-phase libraries of synthetic ligands based on distinct scaffolds were synthesized by combinatorial chemistry. The libraries comprising ligands representing different hydrophobic/hydrophilic properties and sizes were tested for binding to randomly selected biomolecules (labelled with a fluorophore). Fluorescence microscopy was revealed to be a reliable and reproducible technique for the detection of lead ligands which strongly bound the target biomolecule. Results obtained by fluorescence intensity measurements in a 96-well format were less consistent, mainly due to challenges related with the accurate dispensing of the solid support.

Original languageEnglish
Pages (from-to)2187-2193
Number of pages7
JournalSeparation Science And Technology
Volume45
Issue number15
DOIs
Publication statusPublished - 2010
Event15th Conference on Bipartitioning and Biopurification Series - Uxbridge, United Kingdom
Duration: 14 Jun 200919 Jun 2009

Keywords

  • Affinity ligands
  • Combinatorial chemistry
  • HTS

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