Chemoselective Immobilization of Proteins by Microcontact Printing and Bio-orthogonal Click Reactions
CHEMBIOCHEM, vol.14, no.18, pp.2464-2471, 2013 (SCI-Expanded, Scopus)
- Publication Type: Article / Article
- Volume: 14 Issue: 18
- Publication Date: 2013
- Doi Number: 10.1002/cbic.201300478
- Journal Name: CHEMBIOCHEM
- Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus
- 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
- Open Archive Collection: AVESIS Open Access Collection
- Middle East Technical University Affiliated: No
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.