Importance of Substrate–Particle Repulsion for Protein-Templated Assembly of Metal Nanoparticles

We study the protein-directed assembly of colloidal gold nanoparticles on de novo designed protein nanofiber templates. Using sequential assembly on glass substrates, we attach positively charged gold nanoparticles to protein nanofibers engineered to have a high density of negatively charged surface residues. Using a combination of electron and optical microscopy, we measure the density of particle attachment and characterize binding specificity. By varying nanoparticle size and pH of the solution, we explore the importance of charge-dependent particle–fiber and particle–substrate interactions. We find an inverse correlation between particle size and attachment density to protein nanofibers, attributed to the balance between size-dependent electrostatic particle–fiber attraction and particle–substrate repulsion. We show pH-dependent particle attachment density and binding specificity in relation to the protonation fraction of each assembly layer. Finally, we employ hyperspectral scattering microscopy to draw conclusions about particle density and interparticle spacings of optically observable particle assemblies.