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Automating Quantitation of Glycans with Stable Isotope Labels using Mass Spectrometry

Glycan Identification and Quantitation with Stable Isotope Labels Figure 1: Schematic representation of SimGlycan software automating data analysis of stable isotope labeled glycans by MS e.g., isotopic labelling using 12[C6]-2-AA)/13[C6]-2-AA glycomics workflows (Prien, J.M., et al. (2010)).

September 28, 2018

Recent trends in glycomics research show rapid growth of innovation related to accurate mass spectrometry based quantitative analysis of glycans. Stable isotope labels facilitate accurate identification and subsequent quantification of glycans using LC-MS or ESI-MS workflows. There are two major types of stable isotope labeling approaches reported in literature namely, (1) mass shift e.g., isotopic labeling using 18O/16O-Water or 12[C6]-2-AA)/13[C6]-2-AA, and (2) isobaric tags e.g., aminoxyTMT6, and QUANTITY. The mass shift method employs chemically similar but isotopically differentiated tags to derivatize glycan samples, so that they can be distinguished and quantitatively compared by mass spectrometry. Here, quantitation of glycans in samples is achieved in MS1. While, in isobaric tags methods, quantification relies on reporter ions generated in MS2 or MS3. Unlike proteomics, there are no adequate software tools to exploit the above-mentioned glycomics methods. We have developed SimGlycan software modules to streamline the analysis of data generated by above qualitative and quantitative methods.

SimGlycan facilitates automated data analysis of glycomics workflows with isotopic labelling using 12[C6]-2-AA)/13[C6]-2-AA. This functionality facilitates quantification of identified glycans. Figure 1 shows a schematic representation of the MS-based glycomics workflow, and MS/MS Spectra of (Man)5 (GlcNAc)2 labeled with 2-AA[12C]/2-AA[13C] reported in Prien, J.M., et al. (2010). To test the capability of SimGlycan software’s functionality to identify glycans using MS/MS data of glycans labeled with 2-AA[12C]/2-AA[13C], the MS/MS spectra of N glycan with (Man)5 in the Prient et al, 2010 were manually converted into two individual spectrum lists, and subjected to SimGlycan software for MS/MS database search. The glycan structure (Man)5 (GlcNAc)2 was correctly identified by the program as ranked 1 structure – the structure that was best explained by the observed product ions – for both the spectrum lists.

For detailed workflow, Please Visit: http://premierbiosoft.com/citations/glycomics-posters-technical-application-notes.html

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