Thermo-Biolithography: A Technique for Patterning Nucleic Acids and Proteins
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- 作者：Rohan Fernandes ; Hyunmin Yi ; Li-Qun Wu ; Gary W. Rubloff ; Reza Ghodssi ; William E. Bentley ; and Gregory F. Payne
- 刊名：Langmuir
- 出版年：2004
- 出版时间：February 3, 2004
- 年：2004
- 卷：20
- 期：3
- 页码：906 - 913
- 全文大小：257K
- 年卷期：v.20,no.3(February 3, 2004)
- ISSN：1520-5827
文摘
We describe a "biolithographic" technique in which the unique properties of biopolymeric materials andthe 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 inorganicsurface (glass or silicon wafer). Chitosan's pH-responsive solubility facilitates film deposition, while thenucleophilic properties of this polysaccharide allow simple chemistries or biochemistries to be used tocovalently attach species to the film. The thermally responsive protein gelatin is then cast on top of thechitosan film, and the gelatin gel serves as a sacrificial "thermoresist". Pattern transfer is accomplishedby applying a heated stamp to melt specific regions of the gelatin thermoresist and selectively expose theunderlying chitosan. Finally, molecules are conjugated to the exposed chitosan sublayer and the sacrificialgelatin 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 standardglutaraldehyde coupling chemistries), and a model green fluorescent protein (using tyrosinase-initiatedconjugation). Because gelatin can be applied and removed under mild conditions, sequential thermo-biolithographic steps can be performed without destroying previously patterned biomacromolecules. Thesestudies represent the first step toward exploiting nature's exquisite specificity for lithographic patterning.