Journal article
Encapsulated Hydrogels by E-beam Lithography and Their Use in Enzyme Cascade Reactions
Langmuir, Vol.32(16), pp.4043-4051
04/26/2016
Handle:
https://hdl.handle.net/2376/105228
PMCID: PMC4852853
PMID: 27078573
Abstract
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 μm. 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.
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Details
- Title
- Encapsulated Hydrogels by E-beam Lithography and Their Use in Enzyme Cascade Reactions
- Creators
- Rock J Mancini - Department of Chemistry and Biochemistry and California NanoSystems Institute, University of California, Los Angeles , 607 Charles E. Young Drive East, Los Angeles, California 90095-1569, United StatesSamantha J Paluck - Department of Chemistry and Biochemistry and California NanoSystems Institute, University of California, Los Angeles , 607 Charles E. Young Drive East, Los Angeles, California 90095-1569, United StatesErhan Bat - Department of Chemistry and Biochemistry and California NanoSystems Institute, University of California, Los Angeles , 607 Charles E. Young Drive East, Los Angeles, California 90095-1569, United StatesHeather D Maynard - Department of Chemistry and Biochemistry and California NanoSystems Institute, University of California, Los Angeles , 607 Charles E. Young Drive East, Los Angeles, California 90095-1569, United States
- Publication Details
- Langmuir, Vol.32(16), pp.4043-4051
- Academic Unit
- Chemistry, Department of
- Publisher
- United States
- Grant note
- T32 GM008496 / NIGMS NIH HHS
- Identifiers
- 99900546827501842
- Language
- English
- Resource Type
- Journal article