期刊
MOLECULES
卷 18, 期 8, 页码 9833-9849出版社
MDPI AG
DOI: 10.3390/molecules18089833
关键词
peptide microarray; antibody diagnostics; SPAAC; ADIBO; kinetics; non-specific binding (NSB); blocking reagents
资金
- University of Applied Sciences of Western Switzerland, HES-SO RCSO 'RealTech', project [RT 06-10]
- University of Applied Sciences Western Switzerland, Valais/Wallis in Sion
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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