Advancing label-free screening technologies to enhance drug discovery efficiency

INTRODUCTION: Hit identification is a pivotal yet resource-intensive stage of early drug discovery, where large chemical libraries are screened to uncover compounds with target-specific activity. Traditional fluorescence- and luminescence-based high-throughput assays, while fast and automation-friendly, suffer from label-associated artifacts, limited biological relevance, and signal interference that can compromise data fidelity. These challenges, coupled with the growing scale of screening campaigns, have intensified the need for more robust and physiologically relevant label-free screening strategies. AREAS COVERED: This review highlights the emergence of label-free detection technologies as powerful alternatives for hit identification. By enabling direct measurement of biomolecular interactions or cellular responses without secondary reporters, these modalities reduce false positives, improve assay reliability, and enhance mechanistic insight. The authors also summarize their operating principles, recent applications, and practical considerations, emphasizing how label-free approaches can strengthen screening accuracy and accelerate early drug discovery. EXPERT OPINION: Label-free assays have rapidly advanced, offering real-time measurements, improved physiological relevance, and expanding throughput for early drug discovery. While these methods reduce artifacts and broaden target compatibility, challenges remain in validating biological relevance and managing complex kinetic data. Recent software innovations, including automated kinetic modeling and high-throughput data pipelines, are accelerating analysis and enhancing scalability.