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Chemoselective Immobilization of Proteins by Microcontact Printing and Bio‐orthogonal Click Reactions
Author(s) -
Tolstyka Zachary P.,
Richardson Wade,
Bat Erhan,
Stevens Caitlin J.,
Parra Dayanara P.,
Dozier Jonathan K.,
Distefano Mark D.,
Dunn Bruce,
Maynard Heather D.
Publication year - 2013
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201300478
Subject(s) - microcontact printing , click chemistry , azide , chemistry , alkyne , combinatorial chemistry , fluorescence , nanotechnology , polymer chemistry , materials science , organic chemistry , catalysis , physics , quantum mechanics
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 proteins—horse heart myoglobin (HHMb) containing an N‐terminal α‐oxoamide and a red fluorescent protein (mCherry‐CVIA) with a C‐terminal alkyne—were immobilized by incubation onto respective stamped functionalized alkanethiol patterns. Pattern formation was confirmed by fluorescence microscopy.