High-throughput detection of low abundance sialylated glycoproteins in human serum by TiO2 enrichment and targeted LC-MS/MS analysis: application to a prostate cancer sample set.

Abstract

Glycopeptide enrichment can be a strategy to allow the detection of peptides belonging to low abundance proteins in complex matrixes such as blood serum or plasma. Though several glycopeptide enrichment protocols have shown excellent sensitivities in this respect, few reports have demonstrated the applicability of these methods to relatively large sample cohorts. In this work, a fast protocol based on TiO2 enrichment and highly sensitive mass spectrometric analysis by Selected Reaction Monitoring (SRM) has been applied to a cohort of serum samples from prostate cancer and benign prostatic hyperplasia patients in order to detect low abundance proteins in a single LC-MS/MS analysis in nanoscale format, without immunodepletion or peptide fractionation. A peptide library of over 700 formerly N-glycosylated peptides was created by data dependent analysis. Then, 16 medium to low abundance proteins were selected for detection by single injection LC-MS/MS based on selected-reaction monitoring. Results demonstrated the consistent detection of the low-level proteins under investigation. Following label-free quantification, four proteins (Adipocyte plasma membrane-associated protein, Periostin, Cathepsin D and Lysosome-associated membrane glycoprotein 2) were found significantly increased in prostate cancer sera compared to the control group. Graphical abstract ᅟ.

• Authors: Caterina Gabriele, Francesco Cantiello, Annalisa Nicastri, Fabio Crocerossa, Giorgio Ivan Russo, Antonio Cicione, Mihai D Vartolomei, Matteo Ferro, Giuseppe Morgia, Giuseppe Lucarelli, Giovanni Cuda, Rocco Damiano, Marco Gaspari
• Journal: Analytical and bioanalytical chemistry (Anal Bioanal Chem) Vol. 411 Issue 3 Pg. 755-763 (Jan 2019) ISSN: 1618-2650 [Electronic] Germany
• PMID: 30483857 (Publication Type: Journal Article)

Chemical References

• Glycoproteins
• Peptide Library
• Peptides
• titanium dioxide
• Titanium
• N-Acetylneuraminic Acid

Topics

• Aged
• Chemical Fractionation
• Chromatography, Liquid (methods)
• Cohort Studies
• Glycoproteins (blood, chemistry)
• Glycosylation
• High-Throughput Screening Assays (methods)
• Humans
• Male
• Middle Aged
• N-Acetylneuraminic Acid (chemistry)
• Peptide Library
• Peptides (blood)
• Prostatic Neoplasms (blood)
• Tandem Mass Spectrometry (methods)
• Titanium (chemistry)