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 required | 2mg or 100µL 10mM DMSO solution |
| Test concentration | 10µM |
| The integrity of the monolayer | 3 days |
| Replicates | N=2 |
| Incubation Condition | Incubation in CO2 incubator at 37ºC for 90 min |
| P-gp inhibitor | Cyclosporin |
| Integrity of the monolayer | TEER>200 Ω•cm2 |
| Analysis method | LC-MS/MS |
| Data delivery | Papp, Efflux Ratio, %Recovery |
| Data Calculation | Papp = 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 Interpretation | Efflux>2.0 without Pgp inhibitor, P-gp substrate |
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Other bioanalytical services in vitro ADME tests from Axispharm bioanalytical laboratory:
- Solubility Test
- Microsomal Stability Assay (Mouse, Rat, Dog, and Monkey)
- Hepatocytes Stability Assay
- Caco-2 Permeability Assay
- CYP450 inhibition
- Plasma Protein Binding
- Blood-to-Plasma Ratio
- LogD