Rapid Covalent Immobilization of Proteins by Phenol-Based Photochemical Cross-Linking.
A strategy for photoinduced covalent immobilization of proteins on phenol-functionalized surfaces is described. Under visible light irradiation, the reaction can be completed within seconds at ambient temperature, with high yields in aqueous solution of physiological conditions. Protein immobilization is based on a ruthenium-catalyzed radical cross-linking reaction between proteins and phenol-modified surfaces, and the process has proven mild enough for lipase, Staphylococcus aureus protein A, and streptavidin to preserve their bioactivity. This strategy was successfully applied to antibody immobilization on different material platforms, including agarose beads, cellulose membranes, and glass wafers, thus providing a generic procedure for rapid biomodification of surfaces.
School of Life Science and Biotechnology and ‡School of Physics and Optoelectronic Engineering, Dalian University of Technology , Dalian, 116023, P. R. China.
Jun
J
Ren
School of Life Science and Biotechnology and ‡School of Physics and Optoelectronic Engineering, Dalian University of Technology , Dalian, 116023, P. R. China.
Kaikai
K
Tian
School of Life Science and Biotechnology and ‡School of Physics and Optoelectronic Engineering, Dalian University of Technology , Dalian, 116023, P. R. China.
Lingyun
L
Jia
School of Life Science and Biotechnology and ‡School of Physics and Optoelectronic Engineering, Dalian University of Technology , Dalian, 116023, P. R. China.
Xiuyou
X
Han
School of Life Science and Biotechnology and ‡School of Physics and Optoelectronic Engineering, Dalian University of Technology , Dalian, 116023, P. R. China.
Mingshan
M
Zhao
D055910:Antibodies, Immobilized / Q000737:chemistry; D002384:Catalysis; D003432:Cross-Linking Reagents / Q000737:chemistry*; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D055767:Immobilized Proteins / Q000737:chemistry; D008049:Lipase / Q000737:chemistry / Q000378:metabolism; D019800:Phenol / Q000737:chemistry; D010777:Photochemistry / Q000379:methods*; D011506:Proteins / Q000737:chemistry*; D012428:Ruthenium / Q000737:chemistry; D012685:Sepharose / Q000737:chemistry; D012710:Serum Albumin, Bovine; D017550:Spectroscopy, Fourier Transform Infrared; D013205:Staphylococcal Protein A / Q000737:chemistry / Q000378:metabolism; D019809:Streptavidin / Q000737:chemistry
D055910:Antibodies, Immobilized; D003432:Cross-Linking Reagents; D055767:Immobilized Proteins; D011506:Proteins; D013205:Staphylococcal Protein A; D012710:Serum Albumin, Bovine; D019800:Phenol; D012428:Ruthenium; D012685:Sepharose; D019809:Streptavidin; D008049:Lipase; D016650:Fluorescein-5-isothiocyanate