Using “spectral families” to assess the reproducibility of glycopeptide enrichment: human serum O-glycosylation revisited
  • Metaadatok
Tartalom: http://real.mtak.hu/44947/
Archvum: MTA Knyvtr
Gyjtemny: Status = Published



Type = Article
Cm:
Using “spectral families” to assess the reproducibility of glycopeptide enrichment: human serum O-glycosylation revisited
Ltrehoz:
Pap, A.
Medzihradszky F., Katalin
Darula, Zsuzsanna
Kiad:
Springer Verlag
Dtum:
2017
Tma:
QD01 Analytical chemistry / analitikai kémia
QH3011 Biochemistry / biokémia
RM Therapeutics. Pharmacology / terápia, gyógyszertan
Tartalmi lers:
Growing evidence on the diverse biological roles of extracellular glycosylation as well as the need for quality control of protein pharmaceuticals make glycopeptide analysis both exciting and important again after a long hiatus. High-throughput O-glycosylation studies have to tackle the complexity of glycosylation as well as technical difficulties and, up to now, have yielded only limited results mostly from single enrichment experiments. In this study, we address the technical reproducibility of the characterization of the most prevalent O-glycosylation (mucin-type core 1 structures) in human serum, using a two-step lectin affinity-based workflow. Our results are based on automated glycopeptide identifications from higher-energy C-trap dissociation and electron transfer dissociation MS/MS data. Assignments meeting strict acceptance criteria served as the foundation for generating “spectral families” incorporating low-scoring MS/MS identifications, supported by accurate mass measurements and expected chromatographic retention times. We show that this approach helped to evaluate the reproducibility of the glycopeptide enrichment more reliably and also contributed to the expansion of the glycoform repertoire of already identified glycosylated sequences. The roadblocks hindering more in-depth investigations and quantitative analyses will also be discussed. © 2016 Springer-Verlag Berlin Heidelberg
Nyelv:
angol
Tpus:
Article
PeerReviewed
info:eu-repo/semantics/article
Formtum:
text
Azonost:
Pap, A. and Medzihradszky F., Katalin and Darula, Zsuzsanna (2017) Using “spectral families” to assess the reproducibility of glycopeptide enrichment: human serum O-glycosylation revisited. ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 409. pp. 539-550. ISSN 1618-2642
Kapcsolat:
MTMT:3149830; doi:10.1007/s00216-016-9960-7