TY - JOUR
T1 - Protein immobilization onto various surfaces using a polymer-bound isocyanate
AU - Kang, Hyun Jin
AU - Cha, Eun Ji
AU - Park, Hee Deung
N1 - Funding Information:
This study was supported by the Korea Ministry of Environment as “Projects for Developing Eco-Innovation Technologies ( GT-11-G-02-001-1 )”. We would like to thank Hyomin Ahn at Sehwa High School and Samantha Reuter for proof reading this manuscript, and the anonymous reviewers for their careful comments on the manuscript.
Publisher Copyright:
© 2014 Elsevier B.V. All rights reserved.
PY - 2015/1/1
Y1 - 2015/1/1
N2 - Silane coupling agents have been widely used for immobilizing proteins onto inorganic surfaces. However, the immobilization method using silane coupling agents requires several treatment steps, and its application is limited to only surfaces containing hydroxyl groups. The aim of this study was to develop a novel method to overcome the limitations of the silane-based immobilization method using a polymer-bound isocyanate. Initially, polymer-bound isocyanate was dissolved in organic solvent and then was used to dip-coat inorganic surfaces. Proteins were then immobilized onto the dip-coated surfaces by the formation of urea bonds between the isocyanate groups of the polymer and the amine groups of the protein. The reaction was verified by FT-IR in which NCO stretching peaks disappeared, and CO and NH stretching peaks appeared after immobilization. The immobilization efficiency of the newly developed method was insensitive to reaction temperatures (4-50 °C), but the efficiency increased with reaction time and reached a maximum after 4 h. Furthermore, the method showed comparable immobilization efficiency to the silane-based immobilization method and was applicable to surfaces that cannot form hydroxyl groups. Taken together, the newly developed method provides a simple and efficient platform for immobilizing proteins onto surfaces.
AB - Silane coupling agents have been widely used for immobilizing proteins onto inorganic surfaces. However, the immobilization method using silane coupling agents requires several treatment steps, and its application is limited to only surfaces containing hydroxyl groups. The aim of this study was to develop a novel method to overcome the limitations of the silane-based immobilization method using a polymer-bound isocyanate. Initially, polymer-bound isocyanate was dissolved in organic solvent and then was used to dip-coat inorganic surfaces. Proteins were then immobilized onto the dip-coated surfaces by the formation of urea bonds between the isocyanate groups of the polymer and the amine groups of the protein. The reaction was verified by FT-IR in which NCO stretching peaks disappeared, and CO and NH stretching peaks appeared after immobilization. The immobilization efficiency of the newly developed method was insensitive to reaction temperatures (4-50 °C), but the efficiency increased with reaction time and reached a maximum after 4 h. Furthermore, the method showed comparable immobilization efficiency to the silane-based immobilization method and was applicable to surfaces that cannot form hydroxyl groups. Taken together, the newly developed method provides a simple and efficient platform for immobilizing proteins onto surfaces.
KW - Coating
KW - Polymer bound isocyanate
KW - Protein immobilization
KW - Silane coupling agent
KW - Surface immobilization
UR - http://www.scopus.com/inward/record.url?scp=84920685626&partnerID=8YFLogxK
U2 - 10.1016/j.apsusc.2014.10.117
DO - 10.1016/j.apsusc.2014.10.117
M3 - Article
AN - SCOPUS:84920685626
SN - 0169-4332
VL - 324
SP - 198
EP - 204
JO - Applied Surface Science
JF - Applied Surface Science
ER -