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Please use this identifier to cite or link to this item: http://hdl.handle.net/1942/23226

Title: In situ monitoring and optimization of CuAAC-mediated protein functionalization of biosurfaces
Authors: Vranken, Tom
Steen Redeker, Erik
Miszta, Adam
Billen, Brecht
Hermens, Wim
de Laat, Bas
Adriaensens, Peter
Guedens, Wanda
Cleij, Thomas J.
Issue Date: 2017
Citation: SENSORS AND ACTUATORS B-CHEMICAL, 238, p. 992-1000
Abstract: With the current trend to miniaturize bioactive surfaces to micro-or nanometer scale, the strategy of immobilization becomes more important. Therefore, there is a growing need for protein immobilization techniques that create both stable and homogeneously covered surfaces in a reproducible way. One of the most promising methods to achieve this is the combination of biological receptors with 'click' chemistry, like the Copper catalyzed Alkyne Azide Cycloaddition (CuAAC). This work presents a full optimization of all aspects of the 'click' chemistry reaction between proteins and surfaces in order to create covalently and homogeneously covered biosurfaces. The coupling procedure is monitored by in situ ellipsometry, a unique characterization technique that offers the opportunity to quantify minute amounts of surface coupled protein mass in real-time. The optimization involves the azidification of a solid silicon support, the alkynation of two proteins, Staphylococcus aureus Protein A (SpA) and Maltose Binding Protein (MBP), as well as the development of a highly reproducible CuAAC 'click' coupling protocol. Using the here optimized protocol, active and reproducible biolayers can be created rapidly. The proposed surface biofunctionalization method combined with ellipsometry forms a unique and promising platform towards the development of highly sensitive, accurate biosensors.
Notes: [Vranken, Tom; Redeker, Erik Steen; Billen, Brecht; Adriaensens, Peter; Guedens, Wanda; Cleij, Thomas J.] Hasselt Univ, Inst Mat Res, Organ & Biopolymer Chem, BE-3590 Diepenbeek, Belgium. [Redeker, Erik Steen; Cleij, Thomas J.] Maastricht Univ, Maastricht Sci Programme, NL-6200 MD Maastricht, Netherlands. [Redeker, Erik Steen; Hermens, Wim; de laat, Bas] Synapse BV, NL-6229 EV Maastricht, Netherlands. [Miszta, Adam; de laat, Bas] Cardiovasc Res Inst Maastricht, NL-6229 ER Maastricht, Netherlands.
URI: http://hdl.handle.net/1942/23226
DOI: 10.1016/j.snb.2016.07.071
ISI #: 000388948600120
ISSN: 0925-4005
Category: A1
Type: Journal Contribution
Validation: ecoom, 2018
Appears in Collections: Research publications

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