Development of a Platform to Enable Fully Automated Cross-Titration Experiments

Area of Science:

• * Pharmacology
• * Drug Discovery
• * Biochemistry

Background:

• * High-throughput screening (HTS) and lead optimization require compound potency and mechanism of action (MOA) determination.
• * Traditional MOA assays, like agonist shift, are data-intensive, limiting throughput.
• * Cross-titration experiments are crucial for MOA but are time-consuming.

Purpose of the Study:

• * To enhance the throughput of compound cross-titration experiments for MOA determination.
• * To automate MOA assays within a screening triage workflow.
• * To enable rapid assessment of numerous compounds for G protein-coupled receptor (GPCR) modulation.

Main Methods:

• * Integration of Hewlett Packard's D300 digital dispenser with a robotics platform.
• * Development of an automated process for on-the-fly cross-titration in 1536-well plates.
• * Utilization of in-house software and hardware for compound management, data tracking, and biological assays (IP1 and Ca²⁺ detection).

Main Results:

• * Successful automation of cross-titration for positive and negative allosteric modulators.
• * Application to two distinct GPCR targets using two different assay detection formats.
• * Nearly 100 compounds tested per target, demonstrating a significant increase in assay throughput.

Conclusions:

• * The developed automated system substantially enhances the efficiency of MOA assays.
• * This platform accelerates the process of identifying and optimizing drug candidates.
• * The technology is applicable to various GPCR targets and assay formats, facilitating drug discovery efforts.