Unlocking the diagnostic potential of peptides in lateral flow assays

The urgency for rapidly deployable diagnostic tests became evident during the COVID-19 pandemic. Lateral flow assays (LFAs) remain a fundamental tool in point-of-care testing due to their simplicity, affordability, and visual readout. Traditional antibody-based LFAs rely on recombinant proteins that are costly and time-consuming to produce. In contrast, synthetic peptides that mimic protein epitopes can be rapidly designed, chemically synthesized, and tailored to specific targets, enabling faster assay development. This review highlights the emerging role of peptides as versatile biorecognition elements in LFAs, emphasizing their rational design, oriented immobilization, and conjugation to nanomaterials. Peptides offer distinct advantages in reproducibility, stability, and scalable production compared to biological antibodies, facilitating reliable and low-cost manufacturing for decentralized diagnostics. Moreover, recent developments in phage display, affibody engineering, and peptide mimotopes have expanded their diagnostic utility across various diseases. Key limitations, including affinity optimization, regulatory validation, and commercialization readiness, are critically assessed providing a balanced perspective on the field. Overall, this review provides a comprehensive overview of current advances and emerging directions in peptide-driven LFAs, emphasizing their growing potential to enhance next-generation diagnostic technologies. • Peptides offer stable and tunable alternatives to antibodies in LFAs. • Advances in peptide design enable precise biomarker recognition. • Optimized immobilization improves LFA sensitivity and reproducibility. • Peptide-LFAs are robust, low-cost tools promising for point-of-care diagnostics. • Nanomaterials drive next-generation peptide-based LFAs. • Key challenges and potential future directions of peptide-based LFAs are presented.