title: ADIBO-based "click" chemistry for diagnostic peptide micro-array fabrication: physicochemical and assay characteristics.
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.
authors: Institute of Life Technologies, University of Applied Sciences Western Switzerland, Sion 1950, Switzerland. denis.prim@hevs.ch
authors: Denis
authors: D
authors: Prim
authors: Fabien
authors: F
authors: Rebeaud
authors: Vincent
authors: V
authors: Cosandey
authors: Roger
authors: R
authors: Marti
authors: Philippe
authors: P
authors: Passeraub
authors: Marc E
authors: ME
authors: Pfeifer
mesh_terms: D057930:Click Chemistry / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay; D007118:Immunoassay / Q000379:methods*; D007700:Kinetics; D010455:Peptides / Q000737:chemistry*
chemical_list: D010455:Peptides
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