Through-a-chip partial filling affinity capillary electrophoresis for estimating binding constants of ligands to receptors.
Talanta
; 74(4): 605-12, 2008 Jan 15.
Article
en En
| MEDLINE
| ID: mdl-18371682
ABSTRACT
In this paper, we describe the development of a microfluidic/capillary electrophoresis (CE) technique employing partial filling affinity capillary electrophoresis (PFACE) to estimate binding constants of ligands to receptors using as model systems carbonic anhydrase B (CAB, EC 4.2.1.1) and vancomycin from Streptomyces orientalis. Using multilayer soft lithography (MSL), a microfluidic device (MD) consisting of fluid and control channels is fabricated and fitted with an external capillary column. Multiple flow channels allows for manipulation of a zone of ligand and sample containing receptor and non-interacting standards into the MD and subsequently into the capillary column. Upon electrophoresis the sample components migrate into the zone of ligand where equilibrium is established. Changes in migration time of the receptor are used in the analysis to obtain a value for the binding interaction. The manipulation of small volumes of solution on the MD minimizes the need of time-consuming pipetting steps.
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Receptores de Superficie Celular
/
Electroforesis Capilar
Idioma:
En
Revista:
Talanta
Año:
2008
Tipo del documento:
Article
País de afiliación:
Estados Unidos