Reference : Capillary electrophoresis as a fragment screening tool to cross-validate hits from ch...
Scientific journals : Article
Physical, chemical, mathematical & earth Sciences : Chemistry
Life sciences : Biochemistry, biophysics & molecular biology
Human health sciences : Pharmacy, pharmacology & toxicology
Capillary electrophoresis as a fragment screening tool to cross-validate hits from chromogenic assay: Application to FXIIa
Davoine, Clara mailto [Université de Liège / Université de Namur - ULiège / UNamur > > CIRM / NARILIS > >]
Fillet, Marianne mailto [Université de Liège - ULiège > Département de pharmacie > Analyse des médicaments >]
Pochet, Lionel mailto [Université de Namur - UNamur > Department of Pharmacy > Namur research Institute for Life Sciences - NARILIS > >]
Yes (verified by ORBi)
[en] Fragment-based lead discovery ; Serine proteinase inhibitors ; Capillary electrophoresis ; Chromogenic assay ; Factor XIIa ; False positive
[en] In this study, a partial-filling affinity capillary electrophoresis (pf-ACE) method was developed for the cross-validation of fragment hits revealed by chromogenic factor XIIa (FXIIa) assay. Chromogenic assay produces false positives, mainly due to spectrophotometric interferences and sample purity issues. pf-ACE was selected as counter-screening technology because of its separative character and the fact that the target does not have to be attached or tagged. The effects of protein plug length, applied voltage and composition of the running buffer were examined and optimized. Detection limit in terms of dissociation constant was estimated at 400 μM. The affinity evaluation was performed close to physiological conditions (pH 7.4, ionic strength 0.13 mol L−1) in a poly (ethylene oxide)-coated capillary of 75 μm internal diameter x 33 cm length with an applied voltage of 3 kV. This method uncovered chromogenic assay's false positives due to zinc contamination. Moreover, pf-ACE supported the evaluation of compounds absorbing at 405 nm.
Centre Interdisciplinaire de Recherche sur le Médicament - CIRM ; Namur Research Institute for Life Sciences - NARILIS
F.R.S.-FNRS - Fonds de la Recherche Scientifique
Development of new compounds targeting coagulation factor XIIa using innovative microfluidic assays in the context of fragment-based drug discovery
Researchers ; Professionals ; Students

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