MDCK-MDR1 cell permeability assay is widely used to screen the permeability of compounds. MDCK-MDR1 cells stably transfer and express the MDR1 gene (ABCB1) in MDCK cells, which encodes and expresses the efflux protein P-gp that causes multiple drug resistance.

Like the Caco-2 permeability assay, MDCK-MDR1 permeability assay calculates the Efflux Ratio by measuring the transport from AB and BA in two different directions, so as to determine whether a molecule has active efflux mediated by P-gp.

MDCK-MDR1 permeability assay helps to understand the mechanism of compound efflux and helps to find problems related to drug permeability in the early development stage. MDCK-MDR1 permeability analysis not only helps to predict the intestinal wall permeability of the compound but also helps to predict the ability of the compound to cross the blood-brain barrier.

MDCK-MDR1 Permeability Assay Protocol

Compound required2mg or 100µL 10mM DMSO solution
Test concentration10µM
The integrity of the monolayer3 days
Incubation ConditionIncubation in CO2 incubator at 37ºC for 90 min
P-gp inhibitorCyclosporin
Integrity of the monolayerTEER>200 Ω•cm2
Analysis methodLC-MS/MS
Data deliveryPapp, Efflux Ratio, %Recovery
Data CalculationPapp = CRecx VRec/ (A x t x C0)
Efflux Ratio = Papp (B→A) /Papp (A→B)
Recovery = [(Vrec x Crec) + (Vd x Cd)] / (Vd x C0)
Data InterpretationEfflux>2.0 without Pgp inhibitor, P-gp substrate

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