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Molecules 2013, 18(8), 9833-9849; doi:10.3390/molecules18089833

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

1
Institute of Life Technologies, University of Applied Sciences Western Switzerland, Sion 1950, Switzerland
2
Institute of Chemistry, University of Applied Sciences Western Switzerland, Fribourg 1705, Switzerland
3
Institute of Sciences and Industrial Technologies, University of Applied Sciences Western Switzerland, Geneva 1202, Switzerland
*
Author to whom correspondence should be addressed.
Received: 1 July 2013 / Revised: 25 July 2013 / Accepted: 6 August 2013 / Published: 16 August 2013
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Abstract

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. View Full-Text
Keywords: peptide microarray; antibody diagnostics; SPAAC; ADIBO; kinetics; non-specific binding (NSB); blocking reagents peptide microarray; antibody diagnostics; SPAAC; ADIBO; kinetics; non-specific binding (NSB); blocking reagents
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This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).

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MDPI and ACS Style

Prim, D.; Rebeaud, F.; Cosandey, V.; Marti, R.; Passeraub, P.; Pfeifer, M.E. ADIBO-Based “Click” Chemistry for Diagnostic Peptide Micro-Array Fabrication: Physicochemical and Assay Characteristics. Molecules 2013, 18, 9833-9849.

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