Detailed Information on Publication Record
2021
Toward an automated workflow for the study of plasma protein-drug interactions based on capillary electrophoresis-frontal analysis combined with in-capillary mixing of interacting partners
MICHALCOVÁ, Lenka, Hana NEVÍDALOVÁ and Zdeněk GLATZBasic information
Original name
Toward an automated workflow for the study of plasma protein-drug interactions based on capillary electrophoresis-frontal analysis combined with in-capillary mixing of interacting partners
Authors
MICHALCOVÁ, Lenka (203 Czech Republic, belonging to the institution), Hana NEVÍDALOVÁ (203 Czech Republic, belonging to the institution) and Zdeněk GLATZ (203 Czech Republic, guarantor, belonging to the institution)
Edition
Journal of Chromatography A, Amsterdam, Elsevier, 2021, 0021-9673
Other information
Language
English
Type of outcome
Článek v odborném periodiku
Field of Study
10600 1.6 Biological sciences
Country of publisher
Netherlands
Confidentiality degree
není předmětem státního či obchodního tajemství
References:
Impact factor
Impact factor: 4.601
RIV identification code
RIV/00216224:14310/21:00118782
Organization unit
Faculty of Science
UT WoS
000603564600006
Keywords in English
capillary electrophoresis-frontal analysis; on-line mixing; TDLFP; protein-drug interaction
Tags
Tags
International impact, Reviewed
Změněno: 19/2/2021 21:17, prof. RNDr. Zdeněk Glatz, CSc.
Abstract
V originále
Capillary electrophoresis-frontal analysis (CE-FA) together with mobility shift affinity CE is the most frequently used mode of affinity CE for a study of plasma protein-drug interactions, which is a substantial part of the early stage of drug discovery. Whereas in the classic CE-FA setup the sample is prepared by off-line mixing of the interaction partners in the sample vial outside the CE instrument and after a short incubation period loaded into the capillary and analysed, in this work a new methodological approach has been developed that combines CE-FA with the mixing of interacting partners directly inside the capillary. This combination gives rise to a fully automated and versatile methodology for the characterization of these binding interactions besides a substantial reduction in the amounts of sample compounds used. The minimization of possible experimental errors due to the full involving of sophisticated CE instrument in the injection procedure, mixing and separation instead of manual manipulation is another fundamental benefit. The in-capillary mixing is based on the transverse diffusion of laminar flow profile methodology introduced by Krylov et al. using its multi-zone injection modification presented by Řemínek at al.. Actually, after the method optimization, the alternate introduction of six plugs of drug and six plugs of bovine serum protein in BGE, each injected for 3 s at a pressure of -10 mbar (-1 kPa) into the capillary filled by BGE, was found to be the best injection procedure. The method repeatability calculated as RSDs of plateau highs of bovine serum albumin and propranolol as model sample compounds were better than 3.44 %. Its applicability was finally demonstrated on the determination of apparent binding parameters of bovine serum albumin for basic drugs propranolol and lidocaine and acid drug phenylbutazone. The values obtained by a new on-line CE-FA methodology are in agreement with values estimated by classic off-line CE-FA, as well as with literature data obtained using different techniques.
Links
| GA19-08358S, research and development project |
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