Posttranslational modification (PTM) of proteins with glycan groups to form glycoproteins is a common biological motif. Glycans are involved in a wide variety of biological and physiological processes, including recognition and regulatory functions, cellular communication, immunological defense, gene expression, and cell growth and development. Aberrant glycosylation of proteins is connected with cell immortalization, transformation, and metastasis. Due to its many biological functions, glycosylation is one of the most-studied PTMs of eukaryotic cell proteins.
Thermo Scientific Pierce Glycoprotein Isolation Kits, Concanavalin A (ConA) and Wheat Germ Agglutinin (WGA), enable the isolation of glycoproteins at the protein level and from complex mixtures such as serum, tissue, and cultured cell lysates. These complete kits contain the immobilized lectins, binding and wash buffers, as well as the columns required to process up to 10 mg of total protein.
Reversed-phase separation on Thermo Scientific low flow systems is an integral component of glycoproteomics workflows and combines seamlessly with Thermo Scientific Orbitrap mass spectrometers. The Thermo Scientific Vanquish Neo UHPLC System is the system of choice for glycoproteomics. The Vanquish Neo UHPLC System combines an unrivaled degree of innovation to deliver 24/7 reproducible separations of complex mixtures at maximum performance for a variety of high-sensitivity LC-MS workflows
Today’s cutting-edge research pushes LC-MS to its limits. Obtaining high confidence insights to enable accurate resolution of subtle differences and avoid costly dead-ends is needed faster than ever before. The Thermo Scientific Orbitrap Eclipse Tribrid mass spectrometer surpasses these limits with new innovations that deliver the ultimate flexibility to expand experimental scope, and with built-in intelligence to ensure the highest data quality and confidence. One system provides maximum insights, so you productively go beyond today’s discovery.
The Thermo Scientific Orbitrap Fusion Lumos Tribrid Mass Spectrometer is HRAM-capable and offers multiple dissociation techniques (CID, HCD, ETD, and EThcD) for thorough glycopeptide characterization. The system's intelligent data acquisition strategies simplify workflows while improving duty cycle and dynamic range. The Orbitrap Fusion Lumos MS is also capable of synchronous precursor selection (SPS) MS3 acquisition, supporting multiplexed quantification of glycopeptides from various biological samples. This workflow is only available on tribrid mass spectrometers.
Glycopeptide data interpretation is challenging because it requires both peptide sequencing and characterization of the glycosylation site(s) and overall glycan composition. Thermo Scientific Proteome Discoverer Software (2.0 and higher) offers Byonic™ Software from Protein Metrics as a node within its architecture. Byonic Software uses HCD data to determine peptide sugar composition, while corresponding ETD/EThcD data elucidate the peptide backbone. This results in an accurate portrayal of the peptide sequence, its site(s) of glycosylation, and its glycan composition.
To elucidate glycopeptide localization and composition, a combination of high-resolution accurate-mass (HRAM) mass spectrometry and multiple dissociation techniques is required. Such versatility is needed for thorough glycopeptide characterization. HCD or CID fragmentation provides glycan composition information, while ETD and EThcD is used to sequence the peptide and determine glycosylation sites.
Read how the functionalities of this innovative instrument combine to produce a significant performance improvement for standard glycomics and glycoproteomics experiments. Additionally, the unique tribrid architecture makes a wide array of novel acquisition experiments possible facilitating workflows that were previously inaccessible with previous generation platforms.
High-Field Asymmetric waveform Ion Mobility Spectrometry (FAIMS) can be used to optimize ion selectivity and reduce chemical backgrounds during mass spectrometry assays. FAIMS selectivity successfully prevents interferences from co-eluting compounds and increases signal to noise ratios by 100-fold or higher.