biostudies-literatureUnknown21(33)Lu HKing Abdullah University of Science and TechnologyRobust and ultrasensitive biosensing platforms for detecting clinically relevant biomarkers from liquid biopsies are vital for precision diagnostics. However, detecting low-abundance biomarkers such as microRNA typically necessitates complex and costly enzyme-based strategies like PCR or isothermal amplification. Here, a materials-driven approach is leveraged to rationally design stimuli-responsive, signal-amplifying, and graphically-encoded hydrogel microparticles, termed Novabeads, for enzyme-free and fluorescence-based biomarker detection. Novabeads incorporate pH-responsive acrylic acid moieties within a polyethylene glycol diacrylate-based network, enabling significant volume reduction (≈5 fold) upon pH modulation. This stimuli-responsive shrinking, coupled with high bioreceptor loading via thiol-ene click chemistry, enables rapid, enzyme-free optical signal amplification. As a proof-of-concept, fluorescently-labeled peptide nucleic acid (PNA) probes are designed for detecting the cancer biomarker miR-16, via a fluorogenic Förster resonance energy transfer (FRET)-based signal. Novabeads exhibit >30 fold signal enhancement over equivalent conventional hydrogel microparticles, driven by three synergistic mechanisms: increased probe loading (≈2.6 fold), enhanced target capture (≈2.8 fold), and shrinkage-driven amplification (≈5 fold), ultimately leading to over 7 fold reduction in detection limit (28.8 pM; 2.9 fmol), and an expanded linear dynamic range. This rationally designed materials-driven biosensing strategy enables next-generation robust, versatile and enzyme-free biosensors for liquid biopsy diagnostics.Small (Weinheim an der Bergstrasse, Germany)e2503990https://www.ebi.ac.uk/biostudies/studies/S-EPMC12372440biostudies-literatureNovabeads: Stimuli-Responsive Signal-Amplifying Hydrogel Microparticles for Enzymeless Fluorescence-Based Detection of microRNA Biomarkers.PMC12372440Lu HHasan EAlsulaiman DSamman FfalseNovabeads: Stimuli-Responsive Signal-Amplifying Hydrogel Microparticles for Enzymeless Fluorescence-Based Detection of microRNA Biomarkers.Robust and ultrasensitive biosensing platforms for detecting clinically relevant biomarkers from liquid biopsies are vital for precision diagnostics. However, detecting low-abundance biomarkers such as microRNA typically necessitates complex and costly enzyme-based strategies like PCR or isothermal amplification. Here, a materials-driven approach is leveraged to rationally design stimuli-responsive, signal-amplifying, and graphically-encoded hydrogel microparticles, termed Novabeads, for enzyme-free and fluorescence-based biomarker detection. Novabeads incorporate pH-responsive acrylic acid moieties within a polyethylene glycol diacrylate-based network, enabling significant volume reduction (≈5 fold) upon pH modulation. This stimuli-responsive shrinking, coupled with high bioreceptor loading via thiol-ene click chemistry, enables rapid, enzyme-free optical signal amplification. As a proof-of-concept, fluorescently-labeled peptide nucleic acid (PNA) probes are designed for detecting the cancer biomarker miR-16, via a fluorogenic Förster resonance energy transfer (FRET)-based signal. Novabeads exhibit >30 fold signal enhancement over equivalent conventional hydrogel microparticles, driven by three synergistic mechanisms: increased probe loading (≈2.6 fold), enhanced target capture (≈2.8 fold), and shrinkage-driven amplification (≈5 fold), ultimately leading to over 7 fold reduction in detection limit (28.8 pM; 2.9 fmol), and an expanded linear dynamic range. This rationally designed materials-driven biosensing strategy enables next-generation robust, versatile and enzyme-free biosensors for liquid biopsy diagnostics.2025-01-01T00:00:00Z2025 Aug2026-05-08T06:53:36.426Z2026-04-07T23:31:28.183ZS-EPMC123724404055757010.1002/smll.202503990