Sequential Multiplex Analyte Capturing for Phosphoprotein Profiling
2010 (English)In: Molecular & Cellular Proteomics, ISSN 1535-9476, E-ISSN 1535-9484, Vol. 9, no 11, 2474-2481 p.Article in journal (Refereed) Published
Microarray-based sandwich immunoassays can simultaneously detect dozens of proteins. However, their use in quantifying large numbers of proteins is hampered by cross-reactivity and incompatibilities caused by the immunoassays themselves. Sequential multiplex analyte capturing addresses these problems by repeatedly probing the same sample with different sets of antibodycoated, magnetic suspension bead arrays. As a miniaturized immunoassay format, suspension bead array-based assays fulfill the criteria of the ambient analyte theory, and our experiments reveal that the analyte concentrations are not significantly changed. The value of sequential multiplex analyte capturing was demonstrated by probing tumor cell line lysates for the abundance of seven different receptor tyrosine kinases and their degree of phosphorylation and by measuring the complex phosphorylation pattern of the epidermal growth factor receptor in the same sample from the same cavity. Molecular & Cellular Proteomics 9:2474-2481, 2010.
Place, publisher, year, edition, pages
2010. Vol. 9, no 11, 2474-2481 p.
Biochemistry and Molecular Biology
IdentifiersURN: urn:nbn:se:kth:diva-30943DOI: 10.1074/mcp.M110.002709ISI: 000285055900011ScopusID: 2-s2.0-78149301758OAI: oai:DiVA.org:kth-30943DiVA: diva2:404733
QC 201103182011-03-182011-03-072011-03-18Bibliographically approved