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

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2013-12-16
Tolstyka, Zachary P.
Richardson, Wade
Bat, Erhan
Stevens, Caitlin J.
Parra, Dayanara P.
Dozier, Jonathan K.
Distefano, Mark D.
Dunn, Bruce
Maynard, Heather D.
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.

Citation Formats
Z. P. Tolstyka et al., “Chemoselective Immobilization of Proteins by Microcontact Printing and Bio-orthogonal Click Reactions,” CHEMBIOCHEM, vol. 14, no. 18, pp. 2464–2471, 2013, Accessed: 00, 2020. [Online]. Available: https://hdl.handle.net/11511/41903.