Characterizing solute transport across cell layers: Artifact correction and parameter extraction from a simplified...

Area of Science:

• Pharmacology and Drug Development
• Biophysical Chemistry
• Computational Biology

Background:

• Epithelial cell layers on transwell inserts are crucial for early drug development, estimating absorption and bioavailability.
• Automated sampling in these assays increases throughput but can introduce artifacts like residual volume and surface adsorption.
• These artifacts distort concentration time series, impacting downstream pharmacokinetic analysis.

Purpose of the Study:

• To develop a mathematical correction for sampling artifacts in transwell assays.
• To implement a three-compartment model for analyzing solute transport across epithelial barriers.
• To enable more accurate characterization of analyte interactions and compound behavior in vitro.

Main Methods:

• Proposed a mathematical correction method to account for sampling artifacts.
• Developed a three-compartment model to capture membrane diffusion, cellular sequestration, and metabolic loss.
• Demonstrated the method using datasets from transwell epithelial barrier transport assays.

Main Results:

• The mathematical correction effectively addresses systematic artifacts in concentration time series.
• The three-compartment model provides mechanistically meaningful parameters compared to apparent permeability (Papp).
• The approach enhances the accuracy of characterizing analyte interactions with epithelial cell layers.

Conclusions:

• The proposed method improves the precision of automated measurements in transwell assays.
• The three-compartment model offers a more robust analysis of solute transport than conventional methods.
• This approach supports better-informed assessments of drug behavior in vitro transport systems.