Surface-engineered nanobeads for regioselective antibody binding: A robust immunoassay platform leveraging catalytic signal amplification

Abstract

Regulating protein interactions and protein corona formation of nanomaterials is crucial for advancing nanomedicine, where surface engineering of nanomaterials plays a pivotal role in precise control over biological interactions. Here, we present a surface-engineered nanoparticle-based immunoassay platform using carboxyl-enriched polystyrene nanobeads (CEPS) with regioselectively controlled antibody-binding properties. Proteomic analysis and theoretical simulation revealed that CEPS has an enhanced Fc-specific binding affinity for immunoglobulins compared to conventional carboxylated polystyrene beads, with a higher surface carboxyl density critically mediating protein interactions. This regioselective antibody binding with unique Fc-specific affinity eliminates the need for complex surface modifications, streamlining the assay process and broadening the applicability across various immunoassay formats. Additionally, incorporating a palladium catalyst within CEPS enables solvent-triggered on-demand catalytic signal amplification using a leucodye substrate, providing a more stable alternative to enzyme-based methods while significantly enhancing detection sensitivity and stability. The platform demonstrated enhanced performance in detecting clinically relevant biomarkers, including C-reactive protein, interferon-gamma, and the receptor-binding domain of SARS-CoV2, achieving lower detection limits and faster response times compared to conventional enzyme-based ELISA systems. Notably, the CEPS-based assay retained catalytic activity for over 140 days at room temperature, underscoring its potential for reliable, long-term use in diverse diagnostic applications.