Proteomics

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Zirconium(IV)-IMAC for phosphopeptide enrichment in phosphoproteomics

ABSTRACT: IMAC and MOAC magnetic microsphere materials were tested with ug amounts of protein extracts from HepG2/C3A cells. We compared Zirconium (IV) IMAC (Zr-IMAC) magnetic microspheres to commonly used Titanium (IV) IMAC (Ti-IMAC) and TiO2 magnetic microspheres 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. Furthermore, optimized Zr-IMAC and Ti-IMAC magnetic microsphere format were compared to HPLC-based Fe(III)-IMAC for same sample amount (200 ug).

INSTRUMENT(S): TripleTOF 5600, Q Exactive HF

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Hepatocyte, Cell Culture

SUBMITTER: Ignacio Arribas  

LAB HEAD: Ole N. Jensen

PROVIDER: PXD018273 | Pride | 2020-11-26

REPOSITORIES: Pride

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Publications

Zirconium(IV)-IMAC Revisited: Improved Performance and Phosphoproteome Coverage by Magnetic Microparticles for Phosphopeptide Affinity Enrichment.

Arribas Diez Ignacio I   Govender Ireshyn I   Naicker Previn P   Stoychev Stoyan S   Jordaan Justin J   Jensen Ole N ON  

Journal of proteome research 20201123 1

Phosphopeptide enrichment is an essential step in large-scale, quantitative phosphoproteomics 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 TiO<sub>2</sub> magnetic microparticles for phosphopeptide enrichment from simple and complex p  ...[more]