Immobilization of peroxidase glycoprotein on gold electrodes modified with mixed epoxy-boronic acid monolayers
(2002) In Journal of the American Chemical Society 124(43). p.12845-12853- Abstract
- The development of bioelectronic enzyme applications requires the immobilization of active proteins onto solid or colloidal substrates such as gold. Coverage of the gold surface with alkanethiol self-assembled monolayers (SAMS) reduces nonspecific adsorption of proteins and also allows the incorporation onto the surface of ligands with affinity for complementary binding sites on native proteins. We present in this work a strategy for the covalent immobilization of glycosylated proteins previously adsorbed through weak, reversible interactions, on tailored SAMS. Boronic acids, which form cyclic esters with saccharides, are incorporated into SAMS to weakly adsorb the glycoprotein onto the electrode surface through their carbohydrate moiety.... (More)
- The development of bioelectronic enzyme applications requires the immobilization of active proteins onto solid or colloidal substrates such as gold. Coverage of the gold surface with alkanethiol self-assembled monolayers (SAMS) reduces nonspecific adsorption of proteins and also allows the incorporation onto the surface of ligands with affinity for complementary binding sites on native proteins. We present in this work a strategy for the covalent immobilization of glycosylated proteins previously adsorbed through weak, reversible interactions, on tailored SAMS. Boronic acids, which form cyclic esters with saccharides, are incorporated into SAMS to weakly adsorb the glycoprotein onto the electrode surface through their carbohydrate moiety. To prevent protein release from the electrode surface, we combine the affinity motif of boronates with the reactivity of epoxy groups to covalently link the protein to heterofunctional boronateepoxy SAMS. The principle underlying our strategy is the increased immobilization rate achieved by the weak interaction-induced proximity effect between slow reacting oxyrane groups in the SAM and nucleophilic residues from adsorbed proteins, which allows the formation of very stable covalent bonds. This approach is exemplified by the use of phenylboronates-oxyrane mixed monolayers as a reactive support and redox-enzyme horseradish peroxidase as glycoprotein for the preparation of peroxidase electrodes. Quartz crystal microbalance, atomic force microscopy, and electrochemical measurements are used to characterize these enzymatic electrodes. These epoxy-boronate functional monolayers; are versatile, stable interfaces, ready to incorporate glycoproteins by incubation under mild conditions. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/324706
- author
- Abad, JM ; Velez, M ; Santamaria, C ; Guisan, JM ; Matheus, PR ; Vazquez, L ; Gazaryan, I ; Gorton, Lo LU ; Gibson, T and Fernandez, VM
- organization
- publishing date
- 2002
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Journal of the American Chemical Society
- volume
- 124
- issue
- 43
- pages
- 12845 - 12853
- publisher
- The American Chemical Society (ACS)
- external identifiers
-
- wos:000178792400041
- scopus:0037202175
- ISSN
- 1520-5126
- DOI
- 10.1021/ja026658p
- language
- English
- LU publication?
- yes
- additional info
- The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Analytical Chemistry (S/LTH) (011001004)
- id
- d181f8a3-48c7-42d4-b658-22a96956313b (old id 324706)
- date added to LUP
- 2016-04-01 16:17:13
- date last changed
- 2022-04-15 03:26:46
@article{d181f8a3-48c7-42d4-b658-22a96956313b,
abstract = {{The development of bioelectronic enzyme applications requires the immobilization of active proteins onto solid or colloidal substrates such as gold. Coverage of the gold surface with alkanethiol self-assembled monolayers (SAMS) reduces nonspecific adsorption of proteins and also allows the incorporation onto the surface of ligands with affinity for complementary binding sites on native proteins. We present in this work a strategy for the covalent immobilization of glycosylated proteins previously adsorbed through weak, reversible interactions, on tailored SAMS. Boronic acids, which form cyclic esters with saccharides, are incorporated into SAMS to weakly adsorb the glycoprotein onto the electrode surface through their carbohydrate moiety. To prevent protein release from the electrode surface, we combine the affinity motif of boronates with the reactivity of epoxy groups to covalently link the protein to heterofunctional boronateepoxy SAMS. The principle underlying our strategy is the increased immobilization rate achieved by the weak interaction-induced proximity effect between slow reacting oxyrane groups in the SAM and nucleophilic residues from adsorbed proteins, which allows the formation of very stable covalent bonds. This approach is exemplified by the use of phenylboronates-oxyrane mixed monolayers as a reactive support and redox-enzyme horseradish peroxidase as glycoprotein for the preparation of peroxidase electrodes. Quartz crystal microbalance, atomic force microscopy, and electrochemical measurements are used to characterize these enzymatic electrodes. These epoxy-boronate functional monolayers; are versatile, stable interfaces, ready to incorporate glycoproteins by incubation under mild conditions.}},
author = {{Abad, JM and Velez, M and Santamaria, C and Guisan, JM and Matheus, PR and Vazquez, L and Gazaryan, I and Gorton, Lo and Gibson, T and Fernandez, VM}},
issn = {{1520-5126}},
language = {{eng}},
number = {{43}},
pages = {{12845--12853}},
publisher = {{The American Chemical Society (ACS)}},
series = {{Journal of the American Chemical Society}},
title = {{Immobilization of peroxidase glycoprotein on gold electrodes modified with mixed epoxy-boronic acid monolayers}},
url = {{http://dx.doi.org/10.1021/ja026658p}},
doi = {{10.1021/ja026658p}},
volume = {{124}},
year = {{2002}},
}