title: Dip-pen nanolithography-assisted protein crystallization.
abstract: 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 crystallization on lipid array patches is also demonstrated in a microfluidic chip, which opens the way toward high-throughput screening to find suitable nucleation and crystal growth conditions. The results demonstrate the use of DPN in directing and inducing protein crystallization on specific surface locations.
authors.affiliation: Department of Bioengineering Sciences, Vrije Universiteit Brussel , 1050 Brussels, Belgium.
authors.forename: Francesco S
authors.initials: FS
authors.lastname: Ielasi
authors.forename: Michael
authors.initials: M
authors.lastname: Hirtz
authors.forename: Sylwia
authors.initials: S
authors.lastname: Sekula-Neuner
authors.forename: Thomas
authors.initials: T
authors.lastname: Laue
authors.forename: Harald
authors.initials: H
authors.lastname: Fuchs
authors.forename: Ronnie G
authors.initials: RG
authors.lastname: Willaert
mesh_terms: D015060:1,2-Dipalmitoylphosphatidylcholine / Q000737:chemistry; D003460:Crystallization / Q000379:methods*; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation; D036103:Nanotechnology / Q000295:instrumentation; D010316:Particle Size; D019809:Streptavidin / Q000737:chemistry*; D013499:Surface Properties
chemical_list: D015060:1,2-Dipalmitoylphosphatidylcholine; D019809:Streptavidin