Capillary electrophoresis as a fragment screening tool to cross-validate hits from chromogenic assay: Application to FXIIa.
Capillary electrophoresis
Chromogenic assay
Factor XIIa
False positive
Fragment-based lead discovery
Serine proteinase inhibitors
Journal
Talanta
ISSN: 1873-3573
Titre abrégé: Talanta
Pays: Netherlands
ID NLM: 2984816R
Informations de publication
Date de publication:
01 May 2021
01 May 2021
Historique:
received:
11
10
2020
revised:
20
12
2020
accepted:
25
01
2021
entrez:
7
3
2021
pubmed:
8
3
2021
medline:
8
3
2021
Statut:
ppublish
Résumé
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
Identifiants
pubmed: 33676706
pii: S0039-9140(21)00084-9
doi: 10.1016/j.talanta.2021.122163
pii:
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
122163Informations de copyright
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