scholarly journals Rapid and reproducible phosphopeptide enrichment by tandem metal oxide affinity chromatography : application to boron deficiency induced phosphoproteomics

2019 ◽  
Vol 98 (2) ◽  
pp. 370-384 ◽  
Author(s):  
Yanmei Chen ◽  
Wolfgang Hoehenwarter
Keyword(s):  
Metal Oxide ◽  
Affinity Chromatography ◽  
Boron Deficiency ◽  
Phosphopeptide Enrichment ◽  
Metal Oxide Affinity Chromatography

scholarly journals Displacers improve the selectivity of phosphopeptide enrichment by metal oxide affinity chromatography

2017 ◽  
Vol 74 (3) ◽  
pp. 200-207
Author(s):  
Yesenia Herrera ◽  
Sandra Contreras ◽  
Magdalena Hernández ◽  
Laura Álvarez ◽  
Yolanda Mora ◽  
...  
Keyword(s):  
Metal Oxide ◽  
Affinity Chromatography ◽  
Phosphopeptide Enrichment ◽  
Metal Oxide Affinity Chromatography

scholarly journals Displacers improve the selectivity of phosphopeptide enrichment by metal oxide affinity chromatography

2017 ◽  
Vol 74 (3) ◽  
pp. 200-207
Author(s):  
Yesenia Herrera ◽  
Sandra Contreras ◽  
Magdalena Hernández ◽  
Laura Álvarez ◽  
Yolanda Mora ◽  
...  
Keyword(s):  
Metal Oxide ◽  
Affinity Chromatography ◽  
Phosphopeptide Enrichment ◽  
Metal Oxide Affinity Chromatography

Tandem Metal-Oxide Affinity Chromatography for Enhanced Depth of Phosphoproteome Analysis

2013 ◽  
pp. 621-632 ◽  
Author(s):  
Gerold J. M. Beckers ◽  
Wolfgang Hoehenwarter ◽  
Horst Röhrig ◽  
Uwe Conrath ◽  
Wolfram Weckwerth
Keyword(s):  
Metal Oxide ◽  
Affinity Chromatography ◽  
Metal Oxide Affinity Chromatography

scholarly journals Zirconium(IV)-IMAC for phosphopeptide enrichment in phosphoproteomics

2020 ◽  
Author(s):  
Ignacio Arribas Diez ◽  
Ireshyn Govender ◽  
Previn Naicker ◽  
Stoyan Stoychev ◽  
Justin Jordaan ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Affinity Chromatography ◽  
Metal Ion ◽  
Large Scale ◽  
Cell Signalling ◽  
Phosphopeptide Enrichment ◽  
Magnetic Microparticles ◽  
Affinity Enrichment ◽  
Metal Oxide Affinity Chromatography ◽  
Enrichment Techniques

ABSTRACTPhosphopeptide enrichment is an essential step in large-scale, quantitative phosphoproteomics studies by mass spectrometry. Several phosphopeptide affinity enrichment techniques exist, such as Immobilized Metal ion Affinity Chromatography (IMAC) and Metal Oxide Affinity Chromatography (MOAC). We compared Zirconium (IV) IMAC (Zr-IMAC) magnetic microparticles to more commonly used Titanium (IV) IMAC (Ti-IMAC) and TiO2 magnetic microparticles for phosphopeptide enrichment from simple and complex protein samples prior phosphopeptide sequencing and characterization by mass spectrometry (LC-MS/MS). We optimized sample-loading conditions to increase phosphopeptide recovery for Zr-IMAC, Ti-IMAC and TiO2 based workflows. The performance of Zr-IMAC was enhanced by 19-22% to recover up to 5173 phosphopeptides from 200 µg of protein extract from HepG2/C3A cells, making Zr-IMAC the preferred method for phosphopeptide enrichment in this study. Ti-IMAC and TiO2 performance were also optimized to improve phosphopeptide numbers by 28% and 35%, respectively. Furthermore, Zr-IMAC based phosphoproteomics in the magnetic microsphere format identified 23% more phosphopeptides than HPLC-based Fe(III)-IMAC for same sample amount (200 µg), thereby adding 37% more uniquely identified phosphopeptides. We conclude that Zr-IMAC improves phosphoproteome coverage and recommend that this affinity enrichment method should be more widely used in biological and biomedical studies of cell signalling and in the search for disease-biomarkers.

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