What does B/A stand for?
B/A stands for basolateral-to-apical
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The involvement of the transporters can be estimated both in the apical-to-basolateral vs basolateral-to-apical permeabilities as well as by using specific transporter inhibitors, like vanadate. In addition, we can also analyze the ability of the compound to inhibit P-gp transporter by inclusion of known P-gp substrates and monitoring their transport +/- the test compounds (see FDA guidance and EMEA guidance).
The efflux ratio (i. e. Papp(B-A)/Papp(A-B)) is calculated by performing a bidirectional MDR1-MDCK permeability assay where the transport of the compound is measured in the apical to basolateral direction as well as the basolateral to apical direction. If the efflux ratio is greater than or equal to 2 then this indicates drug efflux is occurring. Prazosin, a known P-gp substrate, is screened as a control compound to confirm that the cells are expressing functional P-gp efflux proteins.
Bidirectional transport studies were conducted using our Caco-2 sub clone with high P-gp expression (CLEFF9). HPLC methods were employed to measure drug transport. All digoxin metabolites were P-gp substrates, although digoxin had the greatest efflux ratio. Erythromycin had no effect on the transport of digoxin, maintaining a basolateral to apical efflux ratio of 14. 8, although it did reduce the efflux ratio of dihydrodigoxin and digoxigenin by 34% and 43%, respectively.
An efflux ratio is calculated from the mean apical to basolateral (A-B) Papp data and basolateral to apical.