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Please use this identifier to cite or link to this item: http://hdl.handle.net/1942/21753

Title: Enhanced Biosensor Platforms for Detecting the Atherosclerotic Biomarker VCAM1 Based on Bioconjugation with Uniformly Oriented VCAM1-Targeting Nanobodies
Authors: Ta, Duy Tien
Guedens, Wanda
Vranken, Tom
Vanschoenbeek, Katrijn
Steen Redeker, Erik
Michiels, Luc
Adriaensens, Peter
Issue Date: 2016
Citation: Biosensors, 6 (3), (ART N° 34)
Abstract: Surface bioconjugation of biomolecules has gained enormous attention for developing advanced biomaterials including biosensors. While conventional immobilization (by physisorption or covalent couplings using the functional groups of the endogenous amino acids) usually results in surfaces with low activity, reproducibility and reusability, the application of methods that allow for a covalent and uniformly oriented coupling can circumvent these limitations. In this study, the nanobody targeting Vascular Cell Adhesion Molecule-1 (NbVCAM1), an atherosclerotic biomarker, is engineered with a C-terminal alkyne function via Expressed Protein Ligation (EPL). Conjugation of this nanobody to azidified silicon wafers and Biacore™ C1 sensor chips is achieved via Copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) “click” chemistry to detect VCAM1 binding via ellipsometry and surface plasmon resonance (SPR), respectively. The resulting surfaces, covered with uniformly oriented nanobodies, clearly show an increased antigen binding affinity, sensitivity, detection limit, quantitation limit and reusability as compared to surfaces prepared by random conjugation. These findings demonstrate the added value of a combined EPL and CuAAC approach as it results in strong control over the surface orientation of the nanobodies and an improved detecting power of their targets—a must for the development of advanced miniaturized, multi-biomarker biosensor platforms.
URI: http://hdl.handle.net/1942/21753
DOI: 10.3390/bios6030034
ISI #: 000381858900007
ISSN: 2079-6374
Category: A1
Type: Journal Contribution
Validation: vabb, 2018
Appears in Collections: Research publications

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