Assay development and high-throughput screening of caspases in microfluidic format

Area of Science:

• Biochemistry
• Drug Discovery
• Biotechnology

Background:

• Caspase proteases are key targets in drug discovery.
• Current high-throughput screening (HTS) methods use microtiter plates, but face challenges with smaller well formats.

Purpose of the Study:

• To evaluate microfluidic assay technology as an alternative to microtiter plates for caspase inhibitor screening.
• To compare microfluidic screening with traditional plate-based screening.

Main Methods:

• Fluorogenic screening assays were developed using the Caliper Technologies Labchip platform for caspase isoforms.
• Caspase-3 was used as a model to demonstrate microfluidic assay development and HTS.
• Microfluidic and plate-based screening were compared regarding reagent use, data quality, and operational ease.

Main Results:

• Microfluidic assays successfully screened caspase isoforms using subnanoliter reagent volumes.
• The technology addresses challenges associated with small-volume liquid handling and potential artifacts in microtiter plates.
• Initial results indicate microfluidic screening is a viable and efficient alternative.

Conclusions:

• Microfluidic assay technology provides an effective platform for HTS of caspase inhibitors.
• This technology offers advantages in reagent conservation and overcomes limitations of miniaturized plate-based assays.
• Further adoption of microfluidics can enhance efficiency and reduce costs in drug discovery screening.