Controlled protein embedment onto Au/Ag core-shell nanoparticles for immuno-labeling of nanosilver surface

Difficulties in stable conjugation of biomolecules to nanosilver surfaces have severely limited the use of silver nanostructures in biological applications. Here, we report a facile antibody conjugation onto gold/silver (Au/Ag) core-shell nanoparticles by stable and uniform embedment of an antibody binding protein, protein G, in silver nanoshells. A rigid helical peptide linker with a terminal cysteine residue was fused to protein G. A mixture of the peptide-fused protein G and space-filling free peptide was reacted with gold nanoparticles (AuNPs) to form a protein G-linked peptide layer on the particle surface. Uniform silver nanoshells were successfully formed on these protein G-AuNPs, while stably embedding protein G-linked peptide layers. Protein G specifically targets the Fc region of an antibody and thus affords properly orientated antibodies on the particle surface. Compared to Au nanoparticles of similar size with randomly adsorbed antibodies, the present immuno-labeled Au/Ag core-shell nanoparticles offered nearly 10-fold higher sensitivities for naked-eye detection of surface bound antigens. In addition, small dye molecules that were bonded to the peptide layer on Au nanoparticles exhibited highly enhanced surface-enhanced Raman scattering (SERS) signals upon Ag shell formation. The present strategy provides a simple but efficient way to conjugate antibodies to nanosilver surfaces, which will greatly facilitate wider use of the superior optical properties of silver nanostructures in biological applications.