Chemoselective Immobilization of Proteins by Microcontact Printing and Bio-orthogonal Click Reactions


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Tolstyka Z. P. , Richardson W., Bat E. , Stevens C. J. , Parra D. P. , Dozier J. K. , ...More

CHEMBIOCHEM, vol.14, no.18, pp.2464-2471, 2013 (Journal Indexed in SCI) identifier identifier identifier

  • Publication Type: Article / Article
  • Volume: 14 Issue: 18
  • Publication Date: 2013
  • Doi Number: 10.1002/cbic.201300478
  • Title of Journal : CHEMBIOCHEM
  • Page Numbers: pp.2464-2471
  • Keywords: chemoselectivity, click chemistry, microcontact printing, protein immobilization, self-assembly, SELF-ASSEMBLED MONOLAYERS, OLIGO(ETHYLENE GLYCOL), SURFACE IMMOBILIZATION, FLUORESCENT PROTEIN, CHEMISTRY, FABRICATION, COVALENT, ADSORPTION, CARBOHYDRATE, NANOSCALE

Abstract

Herein, a combination of microcontact printing of functionalized alkanethiols and site-specific modification of proteins is utilized to chemoselectively immobilize proteins onto gold surfaces, either by oxime- or copper-catalyzed alkyne-azide click chemistry. Two molecules capable of click reactions were synthesized, an aminooxy-functionalized alkanethiol and an azide-functionalized alkanethiol, and self-assembled monolayer (SAM) formation on gold was confirmed by IR spectroscopy. The alkanethiols were then individually patterned onto gold surfaces by microcontact printing. Site-specifically modified proteinshorse heart myoglobin (HHMb) containing an N-terminal -oxoamide and a red fluorescent protein (mCherry-CVIA) with a C-terminal alkynewere immobilized by incubation onto respective stamped functionalized alkanethiol patterns. Pattern formation was confirmed by fluorescence microscopy.