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Profiling Glycoproteins Enriched by Multinanoparticle Protein Corona.

Emmajay Sutherland1, Tim S Veth1, Kayla A Markuson1

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This summary is machine-generated.

Nanoparticle enrichment improves analysis of glycoproteins in biofluids like plasma and cerebrospinal fluid (CSF). Combining nanoparticle and MAX enrichment enhances N-glycoproteome coverage, while nanoparticle enrichment alone is better for O-glycoproteome analysis.

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Area of Science:

  • Biochemistry
  • Proteomics
  • Glycomics

Background:

  • Biofluid analysis faces challenges due to complex protein composition and wide dynamic range.
  • Nanoparticle (NP)-based enrichment methods show promise for improving proteomic sampling depth.
  • The impact of NP enrichment on glycoproteome characterization is underexplored.

Purpose of the Study:

  • To investigate nanoparticle (NP) enrichment with and without mixed-mode anion exchange (MAX) for glycopeptide characterization in human plasma and CSF.
  • To assess the impact of NP enrichment on N- and O-linked glycoproteome coverage.
  • To explore the application of NP enrichment for CSF analysis.

Main Methods:

  • Utilized an automated commercial platform for NP enrichment using multinanoparticle protein corona.
  • Applied subsequent MAX enrichment to a subset of samples.
  • Analyzed N- and O-linked glycopeptides in human plasma and CSF.

Main Results:

  • NP enrichment significantly enhanced N-glycoproteome coverage, especially in plasma, and improved detection of low-abundance N-glycopeptides.
  • Combined NP and MAX enrichment further boosted N-glycoproteome improvements.
  • NP enrichment alone was superior for O-glycoproteome analysis, suggesting MAX preferentially enriches N-glycopeptides.
  • First-time application of NP enrichment to CSF revealed increased proteome and glycoproteome depth, including extracellular vesicle biomarkers.

Conclusions:

  • NP enrichment, particularly when combined with MAX, is effective for enhancing N-glycoproteome coverage in human biofluids.
  • NP enrichment alone is optimal for O-glycoproteome analysis.
  • NP enrichment offers a valuable framework for glycoprotein analysis in complex biofluids like CSF, aiding biomarker discovery.