Functional cell-based assays in microliter volumes for ultra-high throughput screening

Area of Science:

• Drug discovery and development
• Cellular and molecular biology
• High-throughput screening technologies

Background:

• Functional cell-based assays are crucial for modern drug discovery, particularly in high-throughput screening (HTS).
• Miniaturized formats, especially 1536-well plates, facilitate screening of large compound libraries (>1 million compounds) at ultra-high throughput (uHTS) rates (>100,000 samples/day).
• Automation and robotics are essential for managing complex assay protocols and ensuring consistent throughput across all screening stages.

Purpose of the Study:

• To review the advancements and feasibility of cell-based assays in miniaturized formats for uHTS.
• To categorize cell-based assays based on response kinetics and readout methods.
• To highlight emerging trends in cell-based uHTS assay development.

Main Methods:

• Categorization of cell-based assays by cellular response kinetics (fast vs. transcriptional).
• Review of readout methodologies for different assay types.
• Discussion of automation and miniaturization strategies in uHTS.

Main Results:

• Cell-based uHTS in microliter volumes is feasible and supported by numerous reports.
• Full automation enables complex assay protocols with multiple liquid handling and detection steps.
• Trends include incorporating assay integral reference signals and kinetic, real-time readouts.

Conclusions:

• Cell-based uHTS is a powerful tool for accelerating drug discovery by enabling large-scale compound screening.
• Incorporating reference signals and kinetic readouts enhances the reliability and informative value of uHTS assays.
• Future cell-based uHTS assays will likely focus on differentiating target hits and elucidating compound mechanisms of action.