Reference : Capillary electrophoretic mobility shift competition assay for the assessment of wea...
Scientific journals : Article
Human health sciences : Pharmacy, pharmacology & toxicology
http://hdl.handle.net/2268/224914
Capillary electrophoretic mobility shift competition assay for the assessment of weak drug-protein interactions
English
Farcas, Elena mailto [Université de Liège - ULiège > Département de pharmacie > Analyse des médicaments >]
Hanson, Julien mailto [Université de Liège - ULiège > Département de pharmacie > Chimie pharmaceutique >]
Pochet, Lionel []
Fillet, Marianne mailto [Université de Liège - ULiège > Département de pharmacie > Analyse des médicaments >]
In press
Analytica Chimica Acta
Elsevier
Yes (verified by ORBi)
International
0003-2670
1873-4324
Netherlands
[en] fragment-based drug discovery ; affinity capillary electrophoresis ; competitive assay ; weak interaction drug-target
[en] Only few reports describe the use of capillary electrophoresis in the context of Fragment Based Drug Discovery (FBDD). In this paper, we will present a generic, fully automated, microscale electrophoretic mobility shift competition assay that can be used in FBDD for primary screening of weak biomolecular interactions between fragments and target protein. The accuracy and reliability of the present method was demonstrated by measuring the interaction between two known fragments inhibiting thrombin, namely benzamidine and p-aminobenzamidine and a relatively weak inhibitor, nafamostat. The measured IC50 were found to be in good accordance with the previously reported ones. Furthermore, we built a small chemical library to evaluate the performance and the advantage of our newly developed screening-bioassay compared to the direct affinity capillary electrophoresis-binding assay. The results demonstrate the high discriminatory power of the method and above all its ability to screen neutral, negatively or positively charged molecules, as well as molecules that have no or low UV-VIS absorbance, greatly expanding the scope of the assay. Finally, we proved that this approach is able to discriminate between competitive binders and irreversible binders. Altogether, this work demonstrates that capillary electrophoresis could constitute an important added value in the arsenal used to screen fragments in drug discovery.
Centre Interdisciplinaire de Recherche sur le Médicament - CIRM
Fonds de la Recherche Scientifique (Communauté française de Belgique) - F.R.S.-FNRS ; Fonds Léon Fredericq
Researchers ; Professionals ; Students
http://hdl.handle.net/2268/224914

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