SERS-Based Immunoassay for α-Fetoprotein Biomarker Detection Using an Au-Ag Nanostars Platform

Spiky Au-Ag nanostars offer intense plasmonic enhancement due to their sharp-tipped morphology, enabling powerful surface-enhanced Raman scattering (SERS). Here, we report a liquid-phase SERS platform that addresses current limitations in cancer biomarker detection, such as low sensitivity and dependence on Raman reporters. Nanostar concentration was tuned by simple centrifugation (10, 30, and 60 min), and their SERS performance was evaluated using methylene blue (MB) and mercaptopropionic acid (MPA) as probe molecules. Signal intensity scaled with nanostar content, enabling sensitive detection. Optimized nanostars were functionalized with MPA, 1-Ethyl-3-(3-dimethylamino1-Ethyl-3-(3dimethylaminopropyl1) carbodiimide (EDC), and N-Hydroxy succinimide (NHS) for covalent attachment of monoclonal anti-α-fetoprotein antibodies (AFP-Ab), facilitating the detection of AFP antigens across 167–38 ng/mL (antibody) and 500–0 ng/mL (antigen) ranges. The limit of detection (LOD) for the antigens was determined to be 16.73 ng/mL. Unlike conventional SERS systems, this aqueous, surfactant-free platform exploits the intrinsic vibrational modes of AFP, enabling sensitive and rapid biomarker detection with strong potential for early cancer diagnostics.