Peptide Tag-Induced Horseradish Peroxidase-Mediated Preparation of a Streptavidin-Immobilized Redox-Sensitive Hydrogel

Several methods have recently been reported for the preparation of redox-sensitive hydrogels using enzymatic reactions, which are useful for encapsulating sensitive materials such as proteins and cells. However, most of the reported hydrogels is difficult to add further function efficiently, limitin...

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Bibliographic Details
Published in:Biomacromolecules 2016-06, Vol.17 (6), p.1978-1984
Main Authors: Mishina, Masahiro, Minamihata, Kosuke, Moriyama, Kousuke, Nagamune, Teruyuki
Format: Article
Language:English
Subjects:
Biopolymers - chemistry
Biopolymers - genetics
Cross-Linking Reagents - chemistry
Cysteine - chemistry
Escherichia coli - genetics
Fibroins - chemistry
Fibroins - genetics
Fibronectins - chemistry
Fibronectins - genetics
Horseradish Peroxidase - chemistry
Hydrogels - chemical synthesis
Hydrogels - chemistry
Oxidation-Reduction
Plasmids
Protein Engineering
Recombinant Fusion Proteins - chemistry
Recombinant Fusion Proteins - genetics
Streptavidin - chemistry
Temperature
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Summary:Several methods have recently been reported for the preparation of redox-sensitive hydrogels using enzymatic reactions, which are useful for encapsulating sensitive materials such as proteins and cells. However, most of the reported hydrogels is difficult to add further function efficiently, limiting the application of the redox-sensitive hydrogels. In this study, peptide sequences of HHHHHHC and GGGGY (Y-tag) were genetically fused to the N- and C-termini of streptavidin (C-SA-Y), respectively, and C-SA-Y was mixed with horseradish peroxidase and thiol-functionalized 4-arm polyethylene glycol to yield a redox-sensitive C-SA-Y immobilized hydrogel (C-SA-Y gel). The C-SA-Y immobilized in the hydrogel retained its affinity for biotin, allowing for the incorporation of proteins and small molecules to hydrogel via biotin. C-SA-Y gel was further prepared within a water-in-oil (w/o) emulsion system to yield a nanosized hydrogel, to which any intracellular and cytotoxic agent can be modified, making it a potential drug delivery carrier.
ISSN:1525-7797
1526-4602
DOI:10.1021/acs.biomac.6b00149