TY - JOUR
T1 - Development of a profiling strategy for metabolic mixtures by combining chromatography and mass spectrometry with cell-based GPCR signaling.
AU - Nijmeijer, S.
AU - Vischer, H.F.
AU - Rudebeck, A.F.
AU - Fleurbaaij, F
AU - Falck, D.
AU - Leurs, R.
AU - Niessen, W.M.A.
AU - Kool, J.
PY - 2012
Y1 - 2012
N2 - In this study, we developed an in-line methodology that combines analytical with pharmacological techniques to characterize metabolites of human histamine H4 receptor (hH4R) ligands. Liquid chromatographic separation of metabolic mixtures is coupled to high-resolution fractionation into 96- or 384-well plates and directly followed by a cell-based reporter gene assay to measure receptor signaling. The complete methodology was designed, optimized, validated, and ultimately miniaturized into a high-density well plate format. Finally, the methodology was demonstrated in a metabolic profiling setting for three hH4R lead compounds and the drug clozapine. This new methodology comprises integrated analytical separations, mass spectrometry, and a cell-based signal transduction-driven reporter gene assay that enables the implementation of comprehensive metabolic profiling earlier in the drug discovery process. © 2012 Society for Laboratory Automation and Screening.
AB - In this study, we developed an in-line methodology that combines analytical with pharmacological techniques to characterize metabolites of human histamine H4 receptor (hH4R) ligands. Liquid chromatographic separation of metabolic mixtures is coupled to high-resolution fractionation into 96- or 384-well plates and directly followed by a cell-based reporter gene assay to measure receptor signaling. The complete methodology was designed, optimized, validated, and ultimately miniaturized into a high-density well plate format. Finally, the methodology was demonstrated in a metabolic profiling setting for three hH4R lead compounds and the drug clozapine. This new methodology comprises integrated analytical separations, mass spectrometry, and a cell-based signal transduction-driven reporter gene assay that enables the implementation of comprehensive metabolic profiling earlier in the drug discovery process. © 2012 Society for Laboratory Automation and Screening.
U2 - 10.1177/1087057112451922
DO - 10.1177/1087057112451922
M3 - Article
SN - 1087-0571
VL - 17
SP - 1329
EP - 1338
JO - Journal of Biomolecular Screening
JF - Journal of Biomolecular Screening
ER -