ADIBO-Based “Click” Chemistry for Diagnostic Peptide Micro-Array Fabrication: Physicochemical and Assay Characteristics

Several azide-derivatized and fluorescently-labeled peptides were immobilized on azadibenzocyclooctyne (ADIBO)-activated slide surfaces via a strain-promoted alkyne-azide cycloaddition (SPAAC) reaction revealing excellent immobilization kinetics, good spot homogeneities and reproducible fluorescence...

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Detalles Bibliográficos
Autores principales: Prim, Denis, Rebeaud, Fabien, Cosandey, Vincent, Marti, Roger, Passeraub, Philippe, Pfeifer, Marc E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2013
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6269721/
https://www.ncbi.nlm.nih.gov/pubmed/23959194
http://dx.doi.org/10.3390/molecules18089833
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author Prim, Denis
Rebeaud, Fabien
Cosandey, Vincent
Marti, Roger
Passeraub, Philippe
Pfeifer, Marc E.
author_facet Prim, Denis
Rebeaud, Fabien
Cosandey, Vincent
Marti, Roger
Passeraub, Philippe
Pfeifer, Marc E.
author_sort Prim, Denis
collection PubMed
description Several azide-derivatized and fluorescently-labeled peptides were immobilized on azadibenzocyclooctyne (ADIBO)-activated slide surfaces via a strain-promoted alkyne-azide cycloaddition (SPAAC) reaction revealing excellent immobilization kinetics, good spot homogeneities and reproducible fluorescence signal intensities. A myc-peptide micro-array immunoassay showed an antibody limit-of-detection (LOD) superior to a microtiter plate-based ELISA. Bovine serum albumin (BSA) and dextran covalently attached via “click” chemistry more efficiently reduced non-specific binding (NSB) of fluorescently-labeled IgG to the microarray surface in comparison to immobilized hexanoic acid and various types of polyethylene glycol (PEG) derivatives. Confirmation of these findings via further studies with other proteins and serum components could open up new possibilities for human sample and microarray platform-based molecular diagnostic tests.
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spelling pubmed-62697212018-12-18 ADIBO-Based “Click” Chemistry for Diagnostic Peptide Micro-Array Fabrication: Physicochemical and Assay Characteristics Prim, Denis Rebeaud, Fabien Cosandey, Vincent Marti, Roger Passeraub, Philippe Pfeifer, Marc E. Molecules Article Several azide-derivatized and fluorescently-labeled peptides were immobilized on azadibenzocyclooctyne (ADIBO)-activated slide surfaces via a strain-promoted alkyne-azide cycloaddition (SPAAC) reaction revealing excellent immobilization kinetics, good spot homogeneities and reproducible fluorescence signal intensities. A myc-peptide micro-array immunoassay showed an antibody limit-of-detection (LOD) superior to a microtiter plate-based ELISA. Bovine serum albumin (BSA) and dextran covalently attached via “click” chemistry more efficiently reduced non-specific binding (NSB) of fluorescently-labeled IgG to the microarray surface in comparison to immobilized hexanoic acid and various types of polyethylene glycol (PEG) derivatives. Confirmation of these findings via further studies with other proteins and serum components could open up new possibilities for human sample and microarray platform-based molecular diagnostic tests. MDPI 2013-08-16 /pmc/articles/PMC6269721/ /pubmed/23959194 http://dx.doi.org/10.3390/molecules18089833 Text en © 2013 by the authors; licensee MDPI, Basel, Switzerland. http://creativecommons.org/licenses/by/3.0/ This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).
spellingShingle Article
Prim, Denis
Rebeaud, Fabien
Cosandey, Vincent
Marti, Roger
Passeraub, Philippe
Pfeifer, Marc E.
ADIBO-Based “Click” Chemistry for Diagnostic Peptide Micro-Array Fabrication: Physicochemical and Assay Characteristics
title ADIBO-Based “Click” Chemistry for Diagnostic Peptide Micro-Array Fabrication: Physicochemical and Assay Characteristics
title_full ADIBO-Based “Click” Chemistry for Diagnostic Peptide Micro-Array Fabrication: Physicochemical and Assay Characteristics
title_fullStr ADIBO-Based “Click” Chemistry for Diagnostic Peptide Micro-Array Fabrication: Physicochemical and Assay Characteristics
title_full_unstemmed ADIBO-Based “Click” Chemistry for Diagnostic Peptide Micro-Array Fabrication: Physicochemical and Assay Characteristics
title_short ADIBO-Based “Click” Chemistry for Diagnostic Peptide Micro-Array Fabrication: Physicochemical and Assay Characteristics
title_sort adibo-based “click” chemistry for diagnostic peptide micro-array fabrication: physicochemical and assay characteristics
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6269721/
https://www.ncbi.nlm.nih.gov/pubmed/23959194
http://dx.doi.org/10.3390/molecules18089833
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