Characterizing solute transport across cell layers : Artifact correction and parameter extraction from a simplified three-compartment model
(2025) In European Journal of Pharmaceutical Sciences 215.- Abstract
Quantifying solute transport across epithelial cell layers grown on transwell inserts is a common approach in early-stage drug development to estimate pharmacokinetic properties such as absorption and bioavailability. To increase throughput and reduce variability, these assays are increasingly automated, including the use of robotic or microfluidic systems for time-resolved sampling. However, both automated and manual sampling can introduce systematic artifacts, such as residual volume retention and surface adsorption, that distort concentration time series and affect downstream analysis. To fully realize the potential precision of automated measurements, we propose a mathematical correction to account for sampling artifacts; then, to... (More)
Quantifying solute transport across epithelial cell layers grown on transwell inserts is a common approach in early-stage drug development to estimate pharmacokinetic properties such as absorption and bioavailability. To increase throughput and reduce variability, these assays are increasingly automated, including the use of robotic or microfluidic systems for time-resolved sampling. However, both automated and manual sampling can introduce systematic artifacts, such as residual volume retention and surface adsorption, that distort concentration time series and affect downstream analysis. To fully realize the potential precision of automated measurements, we propose a mathematical correction to account for sampling artifacts; then, to fit the corrected data to a three-compartment model that captures membrane diffusion, cellular sequestration, and metabolic loss. The method is demonstrated on datasets from transwell epithelial barrier transport assays. We suggest that the considered three-compartment model yields mechanistically more meaningful parameters than the conventional apparent permeability (Papp) measure. The proposed approach thus enables more accurate characterization of analyte interactions with the barrier cell layer, supporting better-informed assessments of compound behavior in vitro transport systems. Quantifying solute transport across epithelial cell layers grown on transwell inserts is a common approach in early-stage drug development to estimate pharmacokinetic properties such as absorption and bioavailability. To increase throughput and reduce variability, these assays are increasingly automated, including the use of robotic or microfluidic systems for time-resolved sampling. However, both automated and manual sampling can introduce systematic artifacts, such as residual volume retention and surface adsorption, that distort concentration time series and affect downstream analysis. To fully realize the potential precision of automated measurements, we propose a mathematical correction to account for sampling artifacts; then, to fit the corrected data to a three-compartment model that captures membrane diffusion, cellular sequestration, and metabolic loss. The method is demonstrated on datasets from transwell epithelial barrier transport assays. We suggest that the considered three-compartment model yields mechanistically more meaningful parameters than the conventional apparent permeability (Papp) measure. The proposed approach thus enables more accurate characterization of analyte interactions with the barrier cell layer, supporting better-informed assessments of compound behavior in vitro transport systems.
(Less)
- author
- Tárnoki-Zách, Júlia ; Boldizsár, Imre ; Kovács, Gábor M. ; Döme, Balázs LU ; Bősze, Szilvia and Czirók, András
- organization
- publishing date
- 2025-12
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Assay interpretation, Modeling, Permeability, Transport kinetics
- in
- European Journal of Pharmaceutical Sciences
- volume
- 215
- article number
- 107323
- publisher
- Elsevier
- external identifiers
-
- pmid:41101515
- scopus:105018706092
- ISSN
- 0928-0987
- DOI
- 10.1016/j.ejps.2025.107323
- language
- English
- LU publication?
- yes
- id
- fcc9a4d2-6088-4511-b9a3-b7107f3132be
- date added to LUP
- 2025-12-11 14:28:09
- date last changed
- 2025-12-11 14:29:28
@article{fcc9a4d2-6088-4511-b9a3-b7107f3132be,
abstract = {{<p>Quantifying solute transport across epithelial cell layers grown on transwell inserts is a common approach in early-stage drug development to estimate pharmacokinetic properties such as absorption and bioavailability. To increase throughput and reduce variability, these assays are increasingly automated, including the use of robotic or microfluidic systems for time-resolved sampling. However, both automated and manual sampling can introduce systematic artifacts, such as residual volume retention and surface adsorption, that distort concentration time series and affect downstream analysis. To fully realize the potential precision of automated measurements, we propose a mathematical correction to account for sampling artifacts; then, to fit the corrected data to a three-compartment model that captures membrane diffusion, cellular sequestration, and metabolic loss. The method is demonstrated on datasets from transwell epithelial barrier transport assays. We suggest that the considered three-compartment model yields mechanistically more meaningful parameters than the conventional apparent permeability (Papp) measure. The proposed approach thus enables more accurate characterization of analyte interactions with the barrier cell layer, supporting better-informed assessments of compound behavior in vitro transport systems. Quantifying solute transport across epithelial cell layers grown on transwell inserts is a common approach in early-stage drug development to estimate pharmacokinetic properties such as absorption and bioavailability. To increase throughput and reduce variability, these assays are increasingly automated, including the use of robotic or microfluidic systems for time-resolved sampling. However, both automated and manual sampling can introduce systematic artifacts, such as residual volume retention and surface adsorption, that distort concentration time series and affect downstream analysis. To fully realize the potential precision of automated measurements, we propose a mathematical correction to account for sampling artifacts; then, to fit the corrected data to a three-compartment model that captures membrane diffusion, cellular sequestration, and metabolic loss. The method is demonstrated on datasets from transwell epithelial barrier transport assays. We suggest that the considered three-compartment model yields mechanistically more meaningful parameters than the conventional apparent permeability (Papp) measure. The proposed approach thus enables more accurate characterization of analyte interactions with the barrier cell layer, supporting better-informed assessments of compound behavior in vitro transport systems.</p>}},
author = {{Tárnoki-Zách, Júlia and Boldizsár, Imre and Kovács, Gábor M. and Döme, Balázs and Bősze, Szilvia and Czirók, András}},
issn = {{0928-0987}},
keywords = {{Assay interpretation; Modeling; Permeability; Transport kinetics}},
language = {{eng}},
publisher = {{Elsevier}},
series = {{European Journal of Pharmaceutical Sciences}},
title = {{Characterizing solute transport across cell layers : Artifact correction and parameter extraction from a simplified three-compartment model}},
url = {{http://dx.doi.org/10.1016/j.ejps.2025.107323}},
doi = {{10.1016/j.ejps.2025.107323}},
volume = {{215}},
year = {{2025}},
}