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

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, an aminooxy-functionalized alkanethiol and an azide-functionalized alkanethiol, were synthesized, 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, respectively were immobilized by incubation onto the stamped functionalized alkanethiol patterns. Pattern formation was confirmed by fluorescence microscopy.

Efficient Functionalization of Additives at Supramolecular Material Surfaces

“…Many examples have been reported using several chemoselective reaction strategies . Covalent postmodification via such a chemoselective reaction is an elegant approach to introduce surface functionality on solid materials.…”

Metal‐Free Click Chemistry Reactions on Surfaces

“…The authors showed that the peptides on the patterned surfaces retained their bioactive function via cell culture studies. More recently, they also reported an efficient and fast immobilization of oxoamide‐modified proteins by microcontact printing onto alkanethiols having alkoxyamino functionalities immobilized on gold surfaces …”

Toward Scalable Flexible Nanomanufacturing for Photonic Structures and Devices

“…Besides thiols, μCP is now widely used in various fields to pattern silanes, biomolecules, proteins, colloidal nanoparticles (NPs), and polymers on metal or semiconductor substrates. SAMs patterned on substrates by μCP induce selective surface dewetting/wetting, which can pattern metals though selective deposition and lift‐off processes .…”