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...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2013-08-01
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| Series: | Molecules |
| Subjects: | |
| Online Access: | http://www.mdpi.com/1420-3049/18/8/9833 |
| _version_ | 1818241417413132288 |
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| author | Marc E. Pfeifer Philippe Passeraub Vincent Cosandey Roger Marti Fabien Rebeaud Denis Prim |
| author_facet | Marc E. Pfeifer Philippe Passeraub Vincent Cosandey Roger Marti Fabien Rebeaud Denis Prim |
| author_sort | Marc E. Pfeifer |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-12T13:29:00Z |
| format | Article |
| id | doaj.art-d6dc54b8b20a4e9d85853edef8b74c03 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-12-12T13:29:00Z |
| publishDate | 2013-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-d6dc54b8b20a4e9d85853edef8b74c032022-12-22T00:23:07ZengMDPI AGMolecules1420-30492013-08-011889833984910.3390/molecules18089833ADIBO-Based “Click” Chemistry for Diagnostic Peptide Micro-Array Fabrication: Physicochemical and Assay CharacteristicsMarc E. PfeiferPhilippe PasseraubVincent CosandeyRoger MartiFabien RebeaudDenis PrimSeveral 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.http://www.mdpi.com/1420-3049/18/8/9833peptide microarrayantibody diagnosticsSPAACADIBOkineticsnon-specific binding (NSB)blocking reagents |
| spellingShingle | Marc E. Pfeifer Philippe Passeraub Vincent Cosandey Roger Marti Fabien Rebeaud Denis Prim ADIBO-Based “Click” Chemistry for Diagnostic Peptide Micro-Array Fabrication: Physicochemical and Assay Characteristics Molecules peptide microarray antibody diagnostics SPAAC ADIBO kinetics non-specific binding (NSB) blocking reagents |
| 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 | peptide microarray antibody diagnostics SPAAC ADIBO kinetics non-specific binding (NSB) blocking reagents |
| url | http://www.mdpi.com/1420-3049/18/8/9833 |
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