Protein addressing on patterned microchip by coupling chitosan electrodeposition and 'electro-click' chemistry.
Many applications in proteomics and lab-on-chip analysis require methods that guide proteins to assemble at surfaces with high spatial and temporal control. Electrical inputs are particularly convenient to control, and there has been considerable effort to discover simple and generic mechanisms that allow electrical inputs to trigger protein assembly on-demand. Here, we report the electroaddressing of a protein to a patterned surface by coupling two generic electroaddressing mechanisms. First, we electrodeposit the stimuli-responsive film-forming aminopolysaccharide chitosan to form a hydrogel matrix at the electrode surface. After deposition, the matrix is chemically functionalized with alkyne groups. Second, we ''electro-click' an azide-tagged protein to the functionalized matrix using electrical signals to trigger conjugation by Huisgen 1,3-dipolar cycloadditions. Specifically, a cathodic potential is applied to the matrix-coated electrode to reduce Cu(II) to Cu(I) which is required for the click reaction. Using fluorescently-labeled bovine serum albumin as our model, we demonstrate that protein conjugation can be controlled spatially and temporally. We anticipate that the coupling of polysaccharide electrodeposition and electro-click chemistry will provide a simple and generic approach to electroaddress proteins within compatible hydrogel matrices.
School of Resource and Environmental Science and Hubei Biomass-Resource Chemistry and Environmental Biotechnology Key Laboratory, Wuhan University, Wuhan, 430079, People's Republic of China.
Xiao-Wen
XW
Shi
Ling
L
Qiu
Zhen
Z
Nie
Ling
L
Xiao
Gregory F
GF
Payne
Yumin
Y
Du
D000480:Alkynes; D001709:Biotechnology; D048271:Chitosan / Q000737:chemistry*; D057930:Click Chemistry / Q000379:methods*; D004595:Electroplating / Q000379:methods*; D055767:Immobilized Proteins / Q000737:chemistry / Q000378:metabolism; D056656:Lab-On-A-Chip Devices; D040901:Proteomics / Q000379:methods*
D000480:Alkynes; D055767:Immobilized Proteins; D048271:Chitosan