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Building a State-of-the-Art Automation Platform to Support High-Throughput 96-Well Caco-2 Permeability Assay.

Xianmei Cai1, Gregory Barker1, Shivani Patel1

  • 1Lead Discovery and Optimization, Bristol Myers Squibb. Co., Princeton, New Jersey, USA.

Assay and Drug Development Technologies
|December 30, 2025
PubMed
Summary
This summary is machine-generated.

This study introduces an automated 96-well Caco-2 permeability assay platform for drug discovery. The system enhances efficiency and capacity for absorption, distribution, metabolism, and excretion profiling, maintaining high predictive accuracy.

Keywords:
96-wellADME profilingCaco-2automationhigh-throughputminiaturizationpermeability assay

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Area of Science:

  • Pharmacology and Drug Discovery
  • Biotechnology and Automation
  • Cell Biology and In Vitro Assays

Background:

  • Automation is vital for efficient in vitro ADME profiling in early drug discovery.
  • Developing automation platforms requires balancing innovation with practical assay needs and technological limits.

Purpose of the Study:

  • To present a state-of-the-art automation system for miniaturizing the Caco-2 permeability assay from 24-well to 96-well format.
  • To demonstrate the efficiency, capacity, and predictive accuracy of the automated platform.

Main Methods:

  • Development of a fully automated 96-well platform integrating cell culture and assay execution infrastructure.
  • Implementation of innovative cleaning protocols, plate tracking, and dynamic scheduling.
  • Validation using 50 literature compounds to assess concordance with the 24-well format.

Main Results:

  • The automated 96-well platform significantly increased efficiency, capacity, and turnaround time for permeability assays.
  • Achieved 94% classification accuracy for literature compounds, showing strong agreement with the 24-well format.
  • Enabled large-scale permeability data generation to support a "predict-first" modeling approach.

Conclusions:

  • The automated 96-well Caco-2 permeability assay platform enhances early drug discovery efficiency and capacity.
  • The platform maintains high predictive accuracy and supports data-driven experimental design.
  • This automation advances the "predict-first" paradigm for compound prioritization and development.