Application of Core–Shell Bimetallic Nanoparticles with Polydopamine-Assisted Nanogap in SERS-Based Lateral Flow Immunoassay of Prolactin

The fabrication of SERS nanotags with efficient antibody loading and high signal enhancement remains a challenging task for combining surface-enhanced Raman spectroscopy (SERS) and lateral flow immunoassay (LFIA). In this study, bimetallic AuDTNB@PDADTNB@Ag nanoparticles with a polydopamine (PDA)-based internal nanogap were synthesized and functionalized with anti-prolactin monoclonal antibodies to produce SERS nanotags. Here, polydopamine serves both as a spacer providing a nanogap between the core and the shell, and as a reaction layer to capture Raman reporter 5,5′-dithiobis(2-nitrobenzoic acid) (DTNB) within the nanogap. Regimes (conditions, protocols) for conjugating antibodies to AuDTNB@PDADTNB@Ag were selected to preserve both the binding affinity for the target analyte and the Raman activity of the SERS nanotag. The SERS nanotag provides plasmonic absorption for visible colorimetric readout, as well as strong SERS signals for highly sensitive quantitative immunoassay. Measuring the Raman intensities of DTNB in the test zone after performing LFIA made it possible to determine prolactin with a detection limit of 0.2 ng/mL in the working range from 1 to 10 ng/mL. The achieved limit of detection was 10-fold lower than the LFIA coupled with colorimetric readout (4.7 ng/mL). The recoveries of prolactin from spiked serum samples were in the range of 70.2–82.6% with relative standard deviations of 2.3–6.8%. Overall, the AuDTNB@PDADTNB@Ag nanotag demonstrated high stability, Raman activity, and specificity, indicating that the SERS nanotag with PDA-assisted internal nanogap is promising for use in SERS immunoassay of other target analytes.