A label-free membrane-based biosensor array with AuNP-modified PDMS for sensitive and specific detection of...

The surface stress-based transduction enabled by antibody functionalization has emerged as a promising paradigm for label-free biosensor development. However, the inherent limitations of conventional detection methods (e.g., high cost, complex modification, insufficient sensitivity) and the challenge of achieving high-throughput analysis hinder their practical clinical application. Herein, a high-throughput label-free biosensor array based on multi-component integration of gold nanoparticles (AuNPs), polydimethylsiloxane (PDMS) membrane, and conductive silver paste is proposed for sensitive and specific detection of alpha-fetoprotein (AFP), a pivotal biomarker for hepatocellular carcinoma (HCC). The specific immunoreaction between immobilized anti-AFP antibodies and target AFP induces compressive surface stress, which triggers PDMS membrane deformation and regulates the electrical resistance of the AuNP conductive layer, realizing the stress-electric coupling transduction. Systematic optimization of antibody surface density (50 μg/mL) and AuNP layer thickness (0.67 μm) maximizes antigen capture efficiency and mechano-electrical transduction performance. The proof-of-concept application for AFP detection demonstrates remarkable analytical performance: a linear detection range of 0-50 ng/mL, a low limit of detection (LOD) of 0.225 ng/mL, high specificity against interfering biomolecules. Furthermore, accurate detection of AFP in clinical serum samples (relative error < 5.05%, RSD < 2.97%) confirms its practical applicability. All these results indicate that the integrated design and parameter optimization of the AuNP-PDMS-based biosensor array enhance the performance of surface stress biosensors, providing a cost-effective platform for the sensitive detection of clinical cancer biomarkers.