Site-specific protein immobilization through N-terminal oxime linkages

Christman, Karen L. and Broyer, Rebecca M. and Tolstyka, Zachary P. and Maynard, Heather D.. (2007) Site-specific protein immobilization through N-terminal oxime linkages. Journal of Materials Chemistry, 17 (19). pp. 2021-2027. ISSN 0959-9428

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Immobilizing proteins in specific orientations is important for diagnostic protein arrays, biomaterials, and other applications where retention of bioactivity is essential. We report an approach for protein micropatterning that exploits a chemoselective reaction to conjugate proteins at the N-terminus to polymer films. A copolymer from 2-hydroxyethyl methacrylate and a Boc-protected aminooxy tetra(ethylene glycol) methacrylate was synthesized by radical polymerization. Boc groups were locally deprotected using photoacid generator-based photolithography. Micropatterns were verified by fluorescence microscopy utilizing green fluorescent aldehyde microspheres. Streptavidin that was subjected to a transamination reaction to install an -ketoamide group at the N-terminus was conjugated to the patterns by oxime bond formation. Since the majority of proteins may be modified to contain a reactive carbonyl group, this methodology should be applicable to pattern a wide variety of proteins specifically through the N-terminus.

Item Type: Article
InterNano Taxonomy: Nanomanufacturing Processes > Self Assembly > Chemical surface functionalization
Nanomanufacturing Processes > Biological Techniques > Protein assembly
Nanoscale Objects and Nanostructured Materials > Nanocomposites > Polymeric
Collections: Nanomanufacturing Research Collection > Nanomanufacturing Nanoscale Science and Engineering Centers > Center for Scalable and Integrated Nanomanufacturing
Depositing User: Moureen Kemei
Date Deposited: 29 Mar 2010 20:11
Last Modified: 30 Sep 2014 15:04

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