期刊
JOURNAL OF SEPARATION SCIENCE
卷 39, 期 21, 页码 4175-4182出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/jssc.201600651
关键词
Affinity chromatography; Gadolinium oxide; Mass spectrometry; Nanoparticles; Phosphopeptides
资金
- Higher Education Commission (HEC) of Pakistan
- Austrian Federal Ministry of Science and Research (BMWF)
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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