Glycobiology Solutions

Glycans, complex carbohydrate molecules, play a critical role in a wide range of biological processes. Understanding their function requires precise analysis of their complex compositions and structures. Mass spectrometry (MS) is increasingly becoming the mainstay technology used to unravel the secrets of glycobiology, enabling researchers to:

• Analyze glycan composition and structure with unrivaled precisions: Identify and characterize glycans with high accuracy.
• Unravel glycan function: Gain insights into how glycans influence the structure and function and affect biological processes of proteins and other molecules to which they are conjugated, such a lipids.
• Advance biomarker discovery: Open up a new era of glycosylation-specific biomarkers for disease prediction, diagnosis, prognosis, and therapy choice.
• Support drug development: Understand the role of glycans as drug targets and as therapeutic molecule modifications.

Focus on Glycoproteomics: Beyond Proteins, Exploring the Glycan World

More than half of all proteins are glycosylated, many of them at multiple sites. Profiling of these glycosylation-related protein isoforms involves techniques to separate, identify, and quantify individual isoforms of glycoproteins and glycopeptides that are present in a biological specimen. Mass spectrometry (MS) is a powerful tool for this purpose. Bruker's solutions, specifically the enablement of high-throughput processing empowered by the glyco-PASEF technology on timsTOF mass spectrometers, offer a cutting-edge approach for glycoprotein and glycan profiling.

Bruker's glyco-PASEF technology, which utilizes the power of time-of-flight (TOF) mass spectrometers equipped with Trapped Ion Mobility Spectrometry (TIMS), offers a revolutionary approach to bottom-up glycoproteomics/peptidomics. This section dives deeper into the specific advantages of this innovative solution for researchers in glycobiology.

TIMS separates ions based on their size and shape (collision cross section, CCS) in addition to their mass-to-charge ratio (m/z). This separation significantly improves: 

• Sensitivity: TIMS reduces ion crowding, allowing for the detection of low-abundance glycopeptides that might be masked in traditional MS analysis.
• Peak Capacity: By separating ions based on additional properties, TIMS resolves complex mixtures of glycopeptides, leading to clearer and more informative spectra.
• Resolution: CCS allows the separation of isomeric/isobaric glycans, i.e. molecules of the same composition and m/z value, but different structures in a uniquely powerful way unparalleled by other approaches. 

Leveraging TIMS' separation capabilities, glyco-PASEF enables the precise targeting of glycopeptides within a sample. This is achieved through:

• Stepped Energy Collision-Induced Dissociation (CID): Differential collsion energy levels are applied to optimally resolve the peptide and glycan components of glycopeptides, with subsequent “addition” of individual spectra to allow a comprehensive analysis.  
• Oxonium-Ion Polygon Gating: TIMS can isolate specific regions of interest within the mass spectrum, by gating on oxonium ions that are emblematic for the presence of glycan moieties.

Glycopeptides can harbor glycans composed of the same monosaccharide building blocks, but with different structural characteristics.

Here, glyco-PASEF and TIMS excel. Mobility-Offset Mass-Aligned (MOMA), based on CCS, leverages the combined power of mass, charge, and mobility data to differentiate between isobaric (same m/z) glycopeptides.

Bruker's GlycoScape™ software streamlines your workflow by providing real-time glycoprotein identification readouts directly from your timsTOF data. This allows researchers to: 

• Make informed decisions:  Gain insights into the data as it's acquired, enabling on-the-fly adjustments to the analysis or prioritization of specific samples.
• Save time: Reduce post-acquisition data analysis time, allowing researchers to focus on interpreting results and drawing conclusions

Bruker’s timsTOF-generated data are agnostic of and have been tested with a number of open source and commercial analytical software products.

• Effective, easy-to-implement interfaces with suites such as MSFraggerGlyco or Protein Metric’s Byonic.
• Allowing the user to work with their preferred glycoproteomic analysis software.