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.
|Number of pages||7|
|Journal||Separation Science And Technology|
|Publication status||Published - 2010|
|Event||15th Conference on Bipartitioning and Biopurification Series - Uxbridge, United Kingdom|
Duration: 14 Jun 2009 → 19 Jun 2009
- Affinity ligands
- Combinatorial chemistry