A method for the design and study of enzyme microstructures formed by means of a flow-through microdispenser
(2001) In Analytical Chemistry 73(17). p.4254-4261- Abstract
- Micrometer-sized enzyme grids were fabricated on gold surfaces using a novel method based on a flow-through microdispenser. The method involves dispensing very small droplets of enzyme solution (similar to 100 pL) during the concomitant relative movement of a gold substrate with respect to the nozzle of a microdispenser, resulting in enzyme patterns with a line width of similar to 100 mum. Different immobilization methods have been evaluated, yielding either enzyme monolayers using functionalized self-assembled thiol monolayers for covalent binding of the enzyme or enzyme multilayers by cross-linking or entrapping the enzymes in a polymer film. The latter immobilization techniques allow the formation of coupled multienzyme structures. On... (More)
- Micrometer-sized enzyme grids were fabricated on gold surfaces using a novel method based on a flow-through microdispenser. The method involves dispensing very small droplets of enzyme solution (similar to 100 pL) during the concomitant relative movement of a gold substrate with respect to the nozzle of a microdispenser, resulting in enzyme patterns with a line width of similar to 100 mum. Different immobilization methods have been evaluated, yielding either enzyme monolayers using functionalized self-assembled thiol monolayers for covalent binding of the enzyme or enzyme multilayers by cross-linking or entrapping the enzymes in a polymer film. The latter immobilization techniques allow the formation of coupled multienzyme structures. On the basis of this feature, coupled bienzyme (glucose oxidase and catalase) or three-enzyme (alpha -glucosidase, mutarotase, and glucose oxidase) microstructures consisting of line patterns of one enzyme intersecting with the patterned lines of the other enzyme(s) were fabricated. By means of scanning electrochemical microscopy (SECM) operated in the generator-collector mode, the enzyme microstructures and their integrity were visualized using the localized detection of enzymatically produced/consumed H2O2. A calibration curve for glucose could be obtained by subsequent SECM line scans over a glucose oxidase microstructure for increasing glucose concentrations, demonstrating the possibility of obtaining localized quantitative data from the prepared microstructures. Possible applications of these enzyme microstructures for multianalyte detection and interference elimination and for screening of different biosensor configurations are highlighted. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/2376335
- author
- Gaspar, S. ; Mosbach, M. ; Wallman, L. ; Laurell, Thomas LU ; Csöregi, Elisabeth LU and Schuhmann, W.
- organization
- publishing date
- 2001
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- SCANNING ELECTROCHEMICAL MICROSCOPY, DIP-PEN NANOLITHOGRAPHY, NANOSTRUCTURES, CHROMATOGRAPHY, FABRICATION, GENERATION, ELECTRODE, DISPENSER, SURFACES
- in
- Analytical Chemistry
- volume
- 73
- issue
- 17
- pages
- 4254 - 4261
- publisher
- The American Chemical Society (ACS)
- external identifiers
-
- wos:000170859300038
- scopus:0035449572
- ISSN
- 1520-6882
- DOI
- 10.1021/ac010214e
- language
- English
- LU publication?
- yes
- id
- 120be11e-0c19-4fad-83e9-91f839d24087 (old id 2376335)
- date added to LUP
- 2016-04-01 12:16:26
- date last changed
- 2022-01-27 01:22:57
@article{120be11e-0c19-4fad-83e9-91f839d24087, abstract = {{Micrometer-sized enzyme grids were fabricated on gold surfaces using a novel method based on a flow-through microdispenser. The method involves dispensing very small droplets of enzyme solution (similar to 100 pL) during the concomitant relative movement of a gold substrate with respect to the nozzle of a microdispenser, resulting in enzyme patterns with a line width of similar to 100 mum. Different immobilization methods have been evaluated, yielding either enzyme monolayers using functionalized self-assembled thiol monolayers for covalent binding of the enzyme or enzyme multilayers by cross-linking or entrapping the enzymes in a polymer film. The latter immobilization techniques allow the formation of coupled multienzyme structures. On the basis of this feature, coupled bienzyme (glucose oxidase and catalase) or three-enzyme (alpha -glucosidase, mutarotase, and glucose oxidase) microstructures consisting of line patterns of one enzyme intersecting with the patterned lines of the other enzyme(s) were fabricated. By means of scanning electrochemical microscopy (SECM) operated in the generator-collector mode, the enzyme microstructures and their integrity were visualized using the localized detection of enzymatically produced/consumed H2O2. A calibration curve for glucose could be obtained by subsequent SECM line scans over a glucose oxidase microstructure for increasing glucose concentrations, demonstrating the possibility of obtaining localized quantitative data from the prepared microstructures. Possible applications of these enzyme microstructures for multianalyte detection and interference elimination and for screening of different biosensor configurations are highlighted.}}, author = {{Gaspar, S. and Mosbach, M. and Wallman, L. and Laurell, Thomas and Csöregi, Elisabeth and Schuhmann, W.}}, issn = {{1520-6882}}, keywords = {{SCANNING ELECTROCHEMICAL MICROSCOPY; DIP-PEN NANOLITHOGRAPHY; NANOSTRUCTURES; CHROMATOGRAPHY; FABRICATION; GENERATION; ELECTRODE; DISPENSER; SURFACES}}, language = {{eng}}, number = {{17}}, pages = {{4254--4261}}, publisher = {{The American Chemical Society (ACS)}}, series = {{Analytical Chemistry}}, title = {{A method for the design and study of enzyme microstructures formed by means of a flow-through microdispenser}}, url = {{http://dx.doi.org/10.1021/ac010214e}}, doi = {{10.1021/ac010214e}}, volume = {{73}}, year = {{2001}}, }