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Encapsulated Hydrogels by E-beam Lithography and Their Use in Enzyme Cascade Reactions
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Date
2016-04-26
Author
Mancini, Rock J.
Paluck, Samantha J.
Bat, Erhan
Maynard, Heather D.
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Electron beam (e-beam) lithography was employed to prepare one protein immobilized hydrogel encapsulated inside another by first fabricating protein-reactive hydrogels of orthogonal reactivity and subsequently conjugating the biomolecules. Exposure of thin films of eight arm star poly(ethylene glycol) (PEG) functionalized with biotin (Biotin PEG), alkyne (Alkyne-PEG) or aminooxy (AO-PEG) end groups to e-beam radiation resulted in cross-linked hydrogels with the respective functionality. It was determined via confocal microscopy that a nominal size exclusion effect exists for streptavidin immobilized on Biotin-PEG hydrogels of feature sizes ranging from 5 to 40 mu m. AO-PEG was subsequently patterned as an encapsulated core inside a contiguous outer shell of Biotin-PEG. Similarly, Alkyne-PEG was patterned as a core inside an AO-PEG shell. The hydrogel reactive end-groups were conjugated to dyes or proteins of complementary reactivity, and the three-dimensional (3-D) spatial orientation was determined for both configurations using confocal microscopy. The enzyme glucose oxidase (GOX) was immobilized in the core of the, encapsulated Alkyne-PEG core/AO-PEG shell architecture, and horseradish peroxidase (HRP) was conjugated to the shell periphery. Bioactivity for the HRP-GOX enzyme pair was observed in this encapsulated configuration by demonstrating that the enzyme pair was capable of enzyme cascade reactions.
Subject Keywords
Spectroscopy
,
Electrochemistry
,
General Materials Science
,
Surfaces and Interfaces
,
Condensed Matter Physics
URI
https://hdl.handle.net/11511/37519
Journal
LANGMUIR
DOI
https://doi.org/10.1021/acs.langmuir.6b00560
Collections
Department of Chemical Engineering, Article
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R. J. Mancini, S. J. Paluck, E. Bat, and H. D. Maynard, “Encapsulated Hydrogels by E-beam Lithography and Their Use in Enzyme Cascade Reactions,”
LANGMUIR
, pp. 4043–4051, 2016, Accessed: 00, 2020. [Online]. Available: https://hdl.handle.net/11511/37519.