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GlycanExplorer™ Workflow

High Throughput Analysis of N-glycans using MS/MS Spectra

GlycanExplorer can efficiently identify glycans using MS/MS data for both native (free reducing end) and derivatized glycan samples generated by direct infusion-, LC-, MALDI- MS/MS workflows. The supported derivatization methods are Permethylation and fluorophore labeled reducing end modification such as 2-aminobenzamide (2-AB), 2-aminobenzoic acid (2-AA), GlycoWorks RapiFluor-MS, Procainamide (ProcA), etc. The software accepts the experimental data and generates candidate peak lists (for LC-MS and MS/MS data), which are further subjected to MS/MS database search to report candidate glycan structures. The search results can be exported, along with glycan structures in a Microsoft Excel file for easy sharing.

The software accommodates three different types of MS/MS database search methods:

  • in-silico Fragment Matching: In this search method, the software reports the candidate glycan structures by matching the experimental product ions with glycan specific in-silico fragments available within the specified target database(s).
  • Spectral Pattern Matching: With the help of built-in or custom created High-Resolution Accurate-Mass (HRAM) glycan spectral libraries, the software allows you to identify probable glycans by comparing the experimental MS/MS spectral pattern with that of the reference spectra available within the target database(s).
  • Combination of in-silico Fragment and Spectral Pattern Matching: GlycanExplorer brings in a unique mechanism of reporting glycan structures that combines the confidence of both in-silico and spectral pattern matching search methods. This workflow identifies the candidate glycans, first by matching the experimental MS/MS fragments with the glycan specific in-silico fragments, then by aligning those experimental spectra (that report a glycan structure with an in-silico score higher than the specified threshold) against the HRAM reference spectral libraries.

Key Features:

1. Import of Thermo Scientific native .raw file format and assign samples and replicates.

2. Efficient LC-MS Peak Detection and Picking.

3. Dedicated window for specifying detailed database search parameters and filters.

4. Facility to perform a database search with multiple adduct combinations in one go.

5. Display the annotated MS/MS spectrum along with the glycan structure.

6. Portable Microsoft Excel report generation.

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Resources

Posters

Introducing a Novel Software Containing an Experimental High-Resolution Mass Spectral Library of N-Glycans...

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Blogs

Spectral Matching Vs. in-silico Fragment Matching – Rendering Confidence to Bio-therapeutic Glycan...

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Webinar

GlycanExplorerTM Software for High Throughput Biotherapeutic Glycan Characterization...

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