A methodology for preparing nanostructured biomolecular interfaces with high enzymatic activity
- 8 December 2011
- journal article
- research article
- Published by Royal Society of Chemistry (RSC) in Nanoscale
- Vol. 4 (2), 659-666
- https://doi.org/10.1039/c1nr11443c
Abstract
The development of a novel method for functionalizing nanopatterned surfaces with catalytically active proteins is reported. This method involves using dip-pen nanolithography (DPN) and polymer pen lithography (PPL) to generate nanoscale patterns of coenzyme A, followed by a phosphopantetheinyl transferase-mediated coupling between coenzyme A and proteins fused to the ybbR-tag. By exploiting the ability to generate protein features over large areas afforded by DPN and PPL, it was now possible to measure protein activity directly on these surfaces. It was found that proteins immobilized on the nanoscale features not only display higher activity per area with decreasing feature size, but are also robust and can be used for repeated catalytic cycles. The immobilization method is applicable to a variety of proteins and gives rise to superior activity compared to proteins attached in random orientations on the surface.Keywords
This publication has 47 references indexed in Scilit:
- Scanning probe block copolymer lithographyProceedings of the National Academy of Sciences, 2010
- Matrix‐Assisted Dip‐Pen Nanolithography and Polymer Pen LithographySmall, 2010
- Dip-pen nanolithography on SiO
x
and tissue-derived substrates: comparison with multiple biological inksScanning, 2009
- Multiplexed Protein Arrays Enabled by Polymer Pen Lithography: Addressing the Inking ChallengeAngewandte Chemie-International Edition, 2009
- Polymer Pen LithographyScience, 2008
- Preparation of $\hbox{F}_{0}\hbox{F}_{1}$ -ATPase Nanoarray by Dip-Pen Nanolithography and Its Application as BiosensorsIEEE Transactions on Nanobioscience, 2008
- Ribosome Display and Dip‐Pen Nanolithography for the Fabrication of Protein NanoarraysAdvanced Materials, 2008
- Direct Site-Selective Covalent Protein Immobilization Catalyzed by a Phosphopantetheinyl TransferaseJournal of the American Chemical Society, 2008
- Electrically Biased Nanolithography with KOH-Coated AFM TipsNano Letters, 2008
- Improved affinity coupling for antibody microarrays: Engineering of double-(His)6-tagged single framework recombinant antibody fragmentsProteomics, 2006