AMP coated SERS NanoTags with hydrophobic locking: Maximizing brightness, stability, and cellular targetability

Developing innovative surface-enhanced Raman scattering (SERS) nanotags continues to attract significant attention due to their unparalleled sensitivity and specificity for in vitro diagnostic and in vivo tumor imaging applications. Here, we report a new class of bright and stable SERS nanotags using alkylmercaptan-PEG (AMP) polymers. Due to its amphiphilic structure and a thiol anchoring group, these polymers strongly absorb onto gold nanoparticles, leading to an inner hydrophobic layer and an outer hydrophilic PEG layer. The inner hydrophobic layer serves to "lock in" the Raman reporter molecules adsorbed on the particle surface via favorable hydrophobic interactions that also allow denser PEG coatings, which "lock out" other molecules from competitive binding or adsorbing to the gold surface, thereby providing superior colloidal and signal stability. The higher grafting densities of AMP polymers compared to conventional thiolated PEG also led to dramatic increases in cellular target selectivity, with specific-to-nonspecific binding ratios reaching beyond an order of magnitude difference. Experimental evaluations and theoretical considerations of dielectric polarization and light scattering indicate that the hydrophobic layer provides a more favorable dielectric environment with less plasmon dampening, greater particle scattering efficiency, and increased Raman reporter polarizability. Accordingly, SERS nanotags with AMP polymer coatings are observed to be considerably brighter (∼10-fold). Furthermore, the AMP-coated SERS nanotag's increased intensity and avidity can boost cellular detection sensitivity by nearly two orders of magnitude.