Microarray-based kinetic colorimetric detection for quantitative multiplex protein phosphorylation analysis

• Verfasst von: Holenya, Pavlo [VerfasserIn]
• Verfasst von: Kitanovic, Igor [VerfasserIn]
• Verfasst von: Heigwer, Florian [VerfasserIn]
• Verfasst von: Wölfl, Stefan [VerfasserIn]
• Titel: Microarray-based kinetic colorimetric detection for quantitative multiplex protein phosphorylation analysis
• Verf.angabe: Pavlo Holenya, Igor Kitanovic, Florian Heigwer, and Stefan Wölfl
• E-Jahr: 2011
• Jahr: 08 March 2011
• Umfang: 5 S.
• Fussnoten: Gesehen am 29.06.2022
• Titel Quelle: Enthalten in: Proteomics
• Ort Quelle: Weinheim : Wiley VCH, 2001
• Jahr Quelle: 2011
• Band/Heft Quelle: 11(2011), 10, Seite 2129-2133
• ISSN Quelle: 1615-9861
• DOI: doi:10.1002/pmic.201000690
• Datenträger: Online-Ressource
• Sprache: eng
• Sach-SW: Antibody microarrays
• Sach-SW: Phosphoproteins
• Sach-SW: Protein arrays
• Sach-SW: Quantitative analysis
• K10plus-PPN: 1808543386

Abstract

Commonly used colorimetric detection applied to protein microarrays with enzymatic signal amplification leads to non-linear signal production upon increase in analyte concentration, thereby considerably limiting the range and accuracy of quantitative readout interpretation. To extend the detection range, we developed a kinetic colorimetric detection protocol for the analysis of ELISA microarrays designed to measure multiple phosphorylated proteins using the platforms ArrayTube™ and ArrayStrip™. With our novel quantification approach, microarrays were calibrated over a broad concentration range spanning four orders of magnitude of analyte concentration with picomolar threshold. We used this design for the simultaneous quantitative measurement of 15 phosphorylated proteins on a single chip.