4.5 Article

Gadolinium oxide: Exclusive selectivity and sensitivity in the enrichment of phosphorylated biomolecules

Journal

JOURNAL OF SEPARATION SCIENCE
Volume 39, Issue 21, Pages 4175-4182

Publisher

WILEY-V C H VERLAG GMBH
DOI: 10.1002/jssc.201600651

Keywords

Affinity chromatography; Gadolinium oxide; Mass spectrometry; Nanoparticles; Phosphopeptides

Funding

  1. Higher Education Commission (HEC) of Pakistan
  2. Austrian Federal Ministry of Science and Research (BMWF)

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Selectivity and sensitivity define the dynamic applicability of separation and enrichment techniques. Owing to proteome complexity, numbers of separation media have been introduced in phosphoproteomics. Complex samples are pretreated to make the low-abundance molecules detectable by mass spectrometry. Gadolinium oxide nanoparticles, offering mono- and bi-dentate interactions, are optimized to capture the phosphopeptides. Selectivity of 1: 11 000 is achieved for digested beta-casein phosphopeptides in bovine serum albumin digest background using gadolinium oxide nanoparticles. The limit of detection goes down to 1 attomole. With the optimized sample preparation protocol, gadolinium oxide nanoparticles enrich phosphopeptides of kappa-casein (Ser(148) and Ser(170)) from digested milk sample, fibrinogen alpha chain phosphopeptide (Ser(609)) along with four hydrolytic products of Ser(22)-modified phosphopeptides from serum.

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