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Enzymatic preparation of streptavidin-immobilized hydrogel using a phenolated linear poly(ethylene glycol)
Institution:1. Department of Applied Chemistry, Graduate School of Engineering, Kyushu University, 744 Motooka, Fukuoka 819-0395, Japan;2. Center for Future Chemistry, Kyushu University, 744 Motooka, Fukuoka 819-0395, Japan;1. Department of Pathology, North Hospital, University Hospital of St-Etienne, St-Etienne, France;2. Department of Pneumonology, North Hospital, University Hospital of St-Etienne, St-Etienne, France;3. Department of Thoracic Surgery, North Hospital, University Hospital of St-Etienne, St-Etienne, France;1. Key Laboratory of Advanced Technique & Preparation for Renewable Energy Materials, Ministry of Education, Yunnan Normal University, Kunming 650500, China;2. Institute of Materials Science and Engineering, Ocean University of China, Qingdao 266100, China;1. Department of Chemical Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, Japan;2. Department of Chemical and Materials Engineering, Tamkang University, Tamsui, New Taipei City 25137, Taiwan;1. Departamento de Química, Universidade Federal de São Carlos, Cx Postal 676, São Carlos 13565-905, São Paulo, Brazil;2. Department of Drug Sciences, University of Pavia, 27100 Pavia, Italy;3. Department of Chemistry, University of Pavia, 27100 Pavia, Italy;4. Instituto Nacional de Ciência e Tecnologia em Tuberculose, Centro de Pesquisas em Biologia Molecular e Funcional, Pontifícia Universidade Católica do Rio Grande do Sul (PUCRS), Porto Alegre, Rio Grande do Sul, Brazil;5. Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto 14040-901, São Paulo, Brazil;1. Department of Molecular Genetics and Cell Biology, The University of Chicago, Chicago, IL 60637, USA;2. Department of Physics, The University of Chicago, Chicago, IL 60637, USA;3. James Franck Institute, The University of Chicago, Chicago, IL 60637, USA;4. Institute for Biophysical Dynamics, The University of Chicago, Chicago, IL 60637, USA;5. Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, IL 60637, USA
Abstract:Hybrid gels constructed from proteins and polymers have attracted a wide range of attention in the field of biomedicine and bioengineering. We report herein the enzymatic preparation of polymer–protein hybrid hydrogels composed of terminally bis-functionalized linear poly(ethylene glycol) (PEG) and streptavidin (SA). PEG was conjugated with tyramine to introduce terminal phenolic hydroxyl (Ph-OH) groups. A peptide tag containing a tyrosine residue (G4Y-tag) was genetically introduced at the C-terminus of SA. The Ph-OH-modified PEG and G4Y-tagged SA (SA-G4Y) were treated by horseradish peroxidase (HRP) in the presence of hydrogen peroxide (H2O2) to yield (PEG-Ph-OH)–(SA-G4Y) hybrid gels. Biotinylated enhanced green fluorescent protein (biotin-EGFP) was selectively captured in the obtained hybrid gels, indicating that SA-G4Y retained its biological function. The amount of biotin-EGFP immobilized in the hybrid gels depended on the concentration of SA-G4Y. In addition, biotinylated bacterial alkaline phosphatase (biotin-BAP) was immobilized in the hybrid gel. The immobilized biotin-BAP exhibited more than 95% of the initial activity after 5 rounds of recycling. The results suggest the facile functionalization of the hybrid gel with a variety of biotinylated functional molecules.
Keywords:Polymer–protein hybrid  Hydrogel  Poly(ethylene glycol)  Streptavidin  Peroxidase  Enzymatic cross-linking
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