Abstract
In this study, protein micropatterns were created on the surface of three-dimensional hydrogel microstructures. Poly(ethylene glycol)(PEG)-based hydrogel microstructures were fabricated on a glass substrate using a poly(dimethylsiloxane) (PDMS) replica as a molding insert and photolithography. The lateral dimension and height of the hydrogel microstructures were easily controlled by the feature size of the photomask and depth of the PDMS replica, respectively. Bovine serum albumin (BSA), a model protein, was covalently immobilized to the surface of the hydrogel microstructure via a 5-azidonitrobenzoyloxy N-hydroxysuccinimide bifunctional linker, which has a phenyl azide group and a protein-binding N-hydroxysuccinimide group on either end. The immobilization of BSA on the PEG hydrogel surface was demonstrated with XPS by confirming the formation of a new nitrogen peak, and the selective immobilization of fluorescent-labeled BSA on the outer region of the three-dimensional hydrogel micropattern was demonstrated by fluorescence. A hydrogel microstructure could immobilize two different enzymes separately, and sequential bienzymatic reaction was demonstrated by reacting glucose and Amplex Red with a hydrogel microstructure where glucose oxidase (GOX) was immobilized on the surface and peroxidase (POD) was encapsulated.
| Original language | English |
|---|---|
| Title of host publication | Responsive Biomaterials for Biomedical Applications |
| Pages | 60-65 |
| Number of pages | 6 |
| Volume | 1095 |
| Publication status | Published - 2008 Dec 1 |
| Event | 2008 MRS Spring Meeting - San Francisco, CA, United States Duration: 2008 Mar 24 → 2008 Mar 28 |
Other
| Other | 2008 MRS Spring Meeting |
|---|---|
| Country | United States |
| City | San Francisco, CA |
| Period | 08/3/24 → 08/3/28 |
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All Science Journal Classification (ASJC) codes
- Mechanical Engineering
- Mechanics of Materials
- Materials Science(all)
- Condensed Matter Physics
Cite this
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Use of hydrogel microstructures as templates for protein immobilization. / Koh, Won G.; Dae, Nylin Kim.
Responsive Biomaterials for Biomedical Applications. Vol. 1095 2008. p. 60-65.Research output: Chapter in Book/Report/Conference proceeding › Conference contribution
TY - GEN
T1 - Use of hydrogel microstructures as templates for protein immobilization
AU - Koh, Won G.
AU - Dae, Nylin Kim
PY - 2008/12/1
Y1 - 2008/12/1
N2 - In this study, protein micropatterns were created on the surface of three-dimensional hydrogel microstructures. Poly(ethylene glycol)(PEG)-based hydrogel microstructures were fabricated on a glass substrate using a poly(dimethylsiloxane) (PDMS) replica as a molding insert and photolithography. The lateral dimension and height of the hydrogel microstructures were easily controlled by the feature size of the photomask and depth of the PDMS replica, respectively. Bovine serum albumin (BSA), a model protein, was covalently immobilized to the surface of the hydrogel microstructure via a 5-azidonitrobenzoyloxy N-hydroxysuccinimide bifunctional linker, which has a phenyl azide group and a protein-binding N-hydroxysuccinimide group on either end. The immobilization of BSA on the PEG hydrogel surface was demonstrated with XPS by confirming the formation of a new nitrogen peak, and the selective immobilization of fluorescent-labeled BSA on the outer region of the three-dimensional hydrogel micropattern was demonstrated by fluorescence. A hydrogel microstructure could immobilize two different enzymes separately, and sequential bienzymatic reaction was demonstrated by reacting glucose and Amplex Red with a hydrogel microstructure where glucose oxidase (GOX) was immobilized on the surface and peroxidase (POD) was encapsulated.
AB - In this study, protein micropatterns were created on the surface of three-dimensional hydrogel microstructures. Poly(ethylene glycol)(PEG)-based hydrogel microstructures were fabricated on a glass substrate using a poly(dimethylsiloxane) (PDMS) replica as a molding insert and photolithography. The lateral dimension and height of the hydrogel microstructures were easily controlled by the feature size of the photomask and depth of the PDMS replica, respectively. Bovine serum albumin (BSA), a model protein, was covalently immobilized to the surface of the hydrogel microstructure via a 5-azidonitrobenzoyloxy N-hydroxysuccinimide bifunctional linker, which has a phenyl azide group and a protein-binding N-hydroxysuccinimide group on either end. The immobilization of BSA on the PEG hydrogel surface was demonstrated with XPS by confirming the formation of a new nitrogen peak, and the selective immobilization of fluorescent-labeled BSA on the outer region of the three-dimensional hydrogel micropattern was demonstrated by fluorescence. A hydrogel microstructure could immobilize two different enzymes separately, and sequential bienzymatic reaction was demonstrated by reacting glucose and Amplex Red with a hydrogel microstructure where glucose oxidase (GOX) was immobilized on the surface and peroxidase (POD) was encapsulated.
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M3 - Conference contribution
SN - 9781605608495
VL - 1095
SP - 60
EP - 65
BT - Responsive Biomaterials for Biomedical Applications
ER -