title: Thermo-biolithography: a technique for patterning nucleic acids and proteins.
abstract: We describe a "biolithographic" technique in which the unique properties of biopolymeric materials and the selective catalytic activities of enzymes are exploited for patterning surfaces under simple and bio-friendly conditions. We begin by coating a reactive film of the polysaccharide chitosan onto an inorganic surface (glass or silicon wafer). Chitosan's pH-responsive solubility facilitates film deposition, while the nucleophilic properties of this polysaccharide allow simple chemistries or biochemistries to be used to covalently attach species to the film. The thermally responsive protein gelatin is then cast on top of the chitosan film, and the gelatin gel serves as a sacrificial "thermoresist". Pattern transfer is accomplished by applying a heated stamp to melt specific regions of the gelatin thermoresist and selectively expose the underlying chitosan. Finally, molecules are conjugated to the exposed chitosan sublayer and the sacrificial gelatin layer is removed (either by treating with warm water or protease). To demonstrate the concept, we patterned a reactive dye (NHS-fluorescein), a model 20-base oligonucleotide (using standard glutaraldehyde coupling chemistries), and a model green fluorescent protein (using tyrosinase-initiated conjugation). Because gelatin can be applied and removed under mild conditions, sequential thermo-biolithographic steps can be performed without destroying previously patterned biomacromolecules. These studies represent the first step toward exploiting nature's exquisite specificity for lithographic patterning.
authors.affiliation: Center for Biosystems Research, University of Maryland Biotechnology Institute, 5115 Plant Sciences Building, College Park, Maryland 20742, USA.
authors.forename: Rohan
authors.initials: R
authors.lastname: Fernandes
authors.forename: Hyunmin
authors.initials: H
authors.lastname: Yi
authors.forename: Li-Qun
authors.initials: LQ
authors.lastname: Wu
authors.forename: Gary W
authors.initials: GW
authors.lastname: Rubloff
authors.forename: Reza
authors.initials: R
authors.lastname: Ghodssi
authors.forename: William E
authors.initials: WE
authors.lastname: Bentley
authors.forename: Gregory F
authors.initials: GF
authors.lastname: Payne
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D048271:Chitosan / Q000737:chemistry; D005452:Fluoresceins / Q000737:chemistry; D005780:Gelatin / Q000737:chemistry; D005821:Genetic Techniques; D049452:Green Fluorescent Proteins / Q000737:chemistry; D009690:Nucleic Acid Conformation; D009693:Nucleic Acid Hybridization; D009696:Nucleic Acids / Q000737:chemistry*; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry*; D013696:Temperature
chemical_list: D005452:Fluoresceins; D009696:Nucleic Acids; D011506:Proteins; D049452:Green Fluorescent Proteins; D005780:Gelatin; D048271:Chitosan