TY - JOUR
T1 - Comparison of fluorescence labelling techniques for the selection of affinity ligands from solid-phase combinatorial libraries
AU - Pina, Ana S.
AU - Lowe, Christopher R.
AU - Roque, Ana Cecília A.
N1 - This work has been financed by Fundac¸ão para a Ciência e Tecnologia (FCT, Portugal) (project PTDC/ BIO/65383/2006), and by British Council-CRUP (Acc¸ão Integrada Luso-Britânica B29/07 and B37/08). Ana Sofia Pina acknowledges a PhD fellowship (SFRH/BD/48804/ 2008) from FCT and a Graduate student bursary from Brunel Graduate School. The authors are also thankful to Dr. Abid Hussain (REQUIMTE, Portugal) and Dr. Jonathan Haigh (Pall Europe) for helpful comments and suggestions.
PY - 2010
Y1 - 2010
N2 - 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.
AB - 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.
KW - Affinity ligands
KW - Combinatorial chemistry
KW - HTS
UR - http://www.scopus.com/inward/record.url?scp=77958610340&partnerID=8YFLogxK
U2 - 10.1080/01496395.2010.507447
DO - 10.1080/01496395.2010.507447
M3 - Conference article
AN - SCOPUS:77958610340
SN - 0149-6395
VL - 45
SP - 2187
EP - 2193
JO - Separation Science And Technology
JF - Separation Science And Technology
IS - 15
T2 - 15th Conference on Bipartitioning and Biopurification Series
Y2 - 14 June 2009 through 19 June 2009
ER -