Glycopeptide Workflow

Site specific glycan analysis

Regulatory agencies have placed increased pressure on biopharmaceutical manufacturers to demonstrate satisfactory programs for understanding, measuring, and controlling glycosylation in glycoprotein-based drugs, with recent updated guidance from the US FDA for characterization of biosimilar therapies.

The Thermo Scientific Orbitrap Eclipse Tribrid Mass Spectrometer offers unique acquisition strategies and workflows through its flexibility in fragmentation types and MSn capabilities that enable the user to go beyond of what is possible in glycan and glycopeptides analyses. Glycan analysis performed at the peptide level provides both glycan composition and peptide sequence at the site of glycosylation. 

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Glycopeptide workflow

glycopeptide workflow

Glycopeptide workflow categories

Digest glycoproteins to glycopeptides in minutes

Obtain high quality analytical results from glycoprotein digests by using Thermo Scientific SMART Digest Kits. A significant advance in sample preparation for biopharmaceutical characterization; SMART Digest Kits utilize immobilized trypsin for fast and simple protein digestion with high reproducibility, high sensitivity, and superior data quality in a format that's compatible with automation.

High resolution reversed-phase glycopeptide separations

Thermo Scientific Acclaim 120 C18 columns are recommended for peptide mapping of biotherapeutic glycoproteins, offering excellent separations and helping to ensure sharp peaks. These columns feature an ultrapure silica substrate with extremely low metal content to minimize tailing effects and deliver symmetrical peak shapes.

Reproducible, easy-to-use UHPLC

The new Thermo Scientific Vanquish Flex UHPLC platform offers the ultimate in biocompatible state-of-the-art binary or quaternary solvent blending. The Vanquish Flex platform is designed to deliver new benchmarks in accuracy, precision, and sensitivity in peptide chromatography.  With rock steady retention times, you can be confident in the identity of every peptide in discovery, development and QC environments.

Seamless compatibility with both DAD, VWD, and mass spectrometric detection and fully compliant Thermo Scientific Chromeleon CDS software makes this the UHPLC system of choice for your entire drug pipeline.

   View the video: Vanquish UHPLC System: Simplifying Biopharma Characterization Workflows
Fragmentation versatility for complete glycopeptide characterization

Determine the exact location of glycosylation with advanced fragmentation techniques on the Thermo Scientific Orbitrap Eclipse Tribrid mass spectrometer. The availability of multiple fragmentation techniques - CID, HCD, ETD; combined ionization spectra such as EThcD; and also product dependent acquisition (such as HCDpdETD), which generates a pair of HCD and ETD spectra. At any stage of MSn, with fragment ion detection in either the ion trap or Orbitrap mass analyzer, offers a new level of versatility and performance for the most challenging applications. Such as, the identification and location of glycopeptides.

   View the video: Orbitrap Fusion Lumos MS: The most sensitive mass spectrometer
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