Surface Functionalization Studies in the Development of Nanohole Plasmonic Sensors

Localized surface plasmon resonance (LSPR) is an optical phenomenon that occurs when light interacts with free electrons on the surface of metallic thin films, producing intensified electromagnetic fields at specific sites, often called “hot spots”. LSPR-based sensing technologies respond to chemical and associated optical interfacial changes. Inherent advantages include enhanced sensitivity, compact size, low production cost, and strong potential for integration into portable, point-of-care diagnostic systems. This study focuses on a detailed investigation into the surface functionalization of localized surface plasmon resonance (LSPR)-based nanohole array (NHA) sensors for biomedical applications. Gold-coated NHA surfaces were functionalized using polyethylene glycol (PEG) self-assembled monolayers (SAMs), enabling specific attachment of biomolecular species. As a proof-of-concept, bovine serum albumin (BSA) and SARS-CoV-2 nanobody proteins were successfully immobilized on the PEGylated surfaces. Individual steps of surface modification including PEGylation, protein immobilization and nanobody immobilization were validated through a dual-method approach which combined measurement of LSPR optical spectral shifts and x-ray photoelectron spectroscopy (XPS) chemical analyses. Reproducibility was assessed across multiple sensors and repeated trials, confirming the repeatability of each functionalization and binding process. The sensor system, consisting of NHA-based plasmonic platform, microfluidics, and a portable optical spectrometer, exhibits the capability for reliable and sensitive, label-free detection of biomolecular targets, including viral antigens, in liquid-phase environments.