A printable hydrogel microarray for drug screening avoids false positives associated with promiscuous aggregating...

Area of Science:

• Biochemistry
• Materials Science
• Drug Discovery

Background:

• High-throughput drug screening often yields false positives from promiscuous inhibitors that form colloidal aggregates.
• These aggregating compounds nonspecifically inhibit enzymes, hindering the identification of viable drug candidates.

Purpose of the Study:

• To develop a printable hydrogel-based platform for accurate differentiation of true enzyme inhibitors from promiscuous aggregating inhibitors.
• To accelerate the drug discovery process by improving the reliability of high-throughput screening.

Main Methods:

• A printable hydrogel platform was developed to immobilize and stabilize enzymes.
• The hydrogel platform was integrated into a microarray format for high-throughput screening.
• Enzyme activity and inhibitor dose-response relationships were quantified using the hydrogel assay.

Main Results:

• The hydrogel platform successfully differentiated true inhibitors from aggregating compounds.
• Enzymes immobilized in hydrogels retained activity despite drying, protease, or chemical denaturant treatment.
• The assay rapidly (<25 min) and inexpensively identified promising lead compounds using 95% less sample than solution assays.

Conclusions:

• The printable hydrogel platform offers a robust solution for identifying true enzyme inhibitors in drug discovery.
• This technology significantly reduces false positives and sample consumption in high-throughput screening.
• The platform accelerates the identification of clinically relevant drug leads with accurate dose-response quantification.