Browsing by Subject "Enzyme immobilization"

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    Stabilization of enzyme-immobilized hydrogels for extended hypoxic cell culture
    (Springer Nature, 2019) Hudson, Britney N.; Dawes, Camron S.; Liu, Hung-Yi; DImmitt, Nathan; Chen, Fangli; Konig, Heiko; Lin, Chien-Chi; Biomedical Engineering, School of Engineering and Technology
    In this work, glucose oxidase (GOx)-immobilized hydrogels are developed and optimized as an easy and convenient means for creating solution hypoxia in a regular incubator. Specifically, acrylated GOx co-polymerizes with poly(ethylene glycol) diacrylate (PEGDA) to form PEGDA-GOx hydrogels. Results show that freeze-drying and reaction by-products, hydrogen peroxide, negatively affect oxygen-consuming activity of network-immobilized GOx. However, the negative effects of freeze-drying can be mitigated by addition of trehalose/raffinose in the hydrogel precursor solution, whereas the inhibition of GOx caused by hydrogen peroxide can be prevented via addition of glutathione (GSH) in the buffer/media. The ability to preserve enzyme activity following freeze-drying and during long-term incubation permits facile application of this material to induce long-term solution/media hypoxia in cell culture plasticware placed in a regular CO2 incubator.