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Plasma Glycopeptide Enrichment

ABSTRACT: We have compared three different glycopeptide enrichment techniques (SAX-ERLIC, HILIC, and lectin affinity) and a C18 control to a tryptic digest of proteins from depleted human plasma.

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Sharon Pitteri

PROVIDER: MSV000082565 | MassIVE | Mon Jul 02 11:49:00 BST 2018

SECONDARY ACCESSION(S): PXD005655

REPOSITORIES: MassIVE

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Publications

Parallel Comparison of N-Linked Glycopeptide Enrichment Techniques Reveals Extensive Glycoproteomic Analysis of Plasma Enabled by SAX-ERLIC.

Totten Sarah M SM Feasley Christa L CL Bermudez Abel A Pitteri Sharon J SJ

Journal of proteome research 20170215 3

Protein glycosylation is of increasing interest due to its important roles in protein function and aberrant expression with disease. Characterizing protein glycosylation remains analytically challenging due to its low abundance, ion suppression issues, and microheterogeneity at glycosylation sites, especially in complex samples such as human plasma. In this study, the utility of three common N-linked glycopeptide enrichment techniques is compared using human plasma. By analysis on an LTQ-Orbitra ...[more]

PMID: 28199111

Similar Datasets

Plasma Glycopeptide Enrichment

Project description:We have compared three different glycopeptide enrichment techniques (SAX-ERLIC, HILIC, and lectin affinity) and a C18 control to a tryptic digest of proteins from depleted human plasma.

2017-02-16 | PXD005655 | Pride

Comparison of Intact Glycopeptide Enrichment Methods Using Strong Anion Exchange and Hydrophilic Interaction Liquid Chromatography

Project description:Heterogeneity of protein glycosylation poses great challenge for analysis that is key to un-puzzle systems glycobiology in diseases. Resolving this conundrum requires global enrichment of glycopeptides for identification and quantitation. To this aim, hydrophilic interaction chromatography (HILIC) has been proved to be an effective approach to enrich N-linked glycopeptides (N-glycopeptides). However, its effectiveness to enrich O-linked glycopeptides (O-glycopeptides) and isobaric labelled glycopeptides remains unclear. Here, we studied three different cartridges in HILIC mode. It was found that removal of N-glycosylation prior to enrichment improved identification of O-glycopeptides. It was noted that increased yield and number of glycosylation identification were seen when using cartridges having materials for strong anion exchange (SAX) chromatography. Remarkably, O-glycopeptides were found to be selectively enriched by HILIC with further improved enrichment being seen when Retain AX cartridge being used. Of particular note, isobaric labelled glycopeptides could be readily enriched by SAX cartridges in HILIC mode to enable quantitative glycoproteomics. It is anticipated that the use of SAX cartridges will facilitate broad applications of qualitative and quantitative glycoproteomics in diseases.

2017-10-19 | PXD005145 | Pride

Evaluation of absorbent cotton for glycopeptide enrichment

Project description:This study introduced the use of cotton in 200µL tips, namely cotton HILIC as a convenient tool for enriching large-scale glycopeptides before MS analysis in this study. The optimal loading buffer for cotton HILIC was investigated by using mouse brain as a complex test sample. Subsequently, the performance of cotton HILIC was evaluated by comparing with other two commercial enrichment approaches, venusil HILIC and Oasis MAX in both mouse brain and seminal plasma. Moreover, capacity and recovery rate were evaluated using different milligram of cotton in 200µL tips. Thus, a simple, convenient, and cost-friendly cotton HILIC method was proposed to achieve highly selective intact glycopeptide profiling.

2023-03-04 | PXD035078 | Pride

A Fully Automated Enrichment and Separation System for Highly-reproducible and In-depth Analysis of Intact Glycopeptide

Project description:A fully automated HILIC-based enrichment and separation system for glycopeptide (AutoGP), which needs no sample collection, was developed in this study.

2024-05-06 | PXD050391 | Pride

Comparison of Intact Glycopeptide Enrichment Methods Using Strong Anion Exchange and Hydrophilic Interaction Liquid Chromatography

2018-07-02 | MSV000082566 | MassIVE

Identifying the glycoproteome of Burkholderia diffusa following knockdown of glycosylation using CRISPR interference - loading control for glycopeptide enrichment

Project description:To interrogate the glycoproteome following knockdown of pglL and subsequent knockdown of glycosylation in Burkholderia diffusa through Zwitterionic Hydrophilic Interaction Liquid Chromatography (ZIC-HILIC) glycopeptide enrichment – loading control pre-enrichment

2023-03-20 | PXD037735 | Pride

Identifying the glycoproteome of Burkholderia thailandensis following knockdown of glycosylation using CRISPR interference - loading control for glycopeptide enrichment

Project description:To interrogate the glycoproteome following knockdown of pglL and subsequent knockdown of glycosylation in Burkholderia thailandensis through Zwitterionic Hydrophilic Interaction Liquid Chromatography (ZIC-HILIC) glycopeptide enrichment – loading control pre-enrichment

2023-03-20 | PXD037748 | Pride

Identifying the glycoproteome of Burkholderia cenocepacia following knockdown of glycosylation using CRISPR interference - loading control for glycopeptide enrichment

2023-03-20 | PXD037749 | Pride

Identifying the glycoproteome of Burkholderia multivorans following knockdown of glycosylation using CRISPR interference - loading control for glycopeptide enrichment

Project description:To interrogate the glycoproteome following knockdown of pglL and subsequent knockdown of glycosylation in Burkholderia multivorans through Zwitterionic Hydrophilic Interaction Liquid Chromatography (ZIC-HILIC) glycopeptide enrichment – loading control pre-enrichment

2023-03-20 | PXD037747 | Pride

Identifying the glycoproteome of Burkholderia cenocepacia following knockdown of glycosylation using CRISPR interference (low dCas9 induction) - loading control for glycopeptide enrichment

Project description:To interrogate the glycoproteome following knockdown of pglL and subsequent knockdown of glycosylation in Burkholderia cenocepacia through Zwitterionic Hydrophilic Interaction Liquid Chromatography (ZIC-HILIC) glycopeptide enrichment with 0.1% rhamnose for dCas9 induction– loading control pre-enrichment. Using strains harbouring the pglL-targeting guide C (Bc-pglL-C) and guideless plasmids.

2023-03-20 | PXD038044 | Pride

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