F. S. Ielasi, M. Hirtz, S. Sekula-Neuner, T. Laue, H. Fuchs, R. G. Willaert 

J. Am. Chem. Soc. 137 (2015) 154-157

We demonstrate the use of dip-pen nanolithography (DPN) to crystallize proteins on surface localized functionalized lipid layer arrays. DOPC lipid layers, containing small amounts of biotin-DOPE lipid molecules, were printed on glass substrates and evaluated in vapor diffusion and batch crystallization screening setups, where streptavidin was used as a model protein for crystallization. Independently of the crystallization system used and the geometry of the lipid layers, nucleation of streptavidin crystals occurred specifically on the DPN-printed biotinylated structures. Protein crys-tallization on lipid array patches is also demonstrat-ed in a microfluidic chip, which opens the way to-wards high-throughput screening to find suitable nucleation and crystal growth conditions. The re-sults demonstrate the use of DPN in directing and inducing protein crystallization on specific surface locations.