MassIVE MSV000083698

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Standardization of PGC-LC-MS-based glycomics for sample specific glycotyping

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Description

This study combined the use of hydrolyzed dextran as an internal standard and Skyline software for post-acquisition normalization to reduce retention time and peak area technical variation in PGC-based glycan analyses. This approach allowed assignment of system-independent retention values that are applicable to typical PGC-based glycan separations and supported the construction of a library containing >300 PGC-separated glycan structures with normalized glucose unit (GU) retention values. The utility of this approach is demonstrated in two ways. First, to inform the search space for bioinformatically predicted but unobserved glycan structures, predictive models for two structural modifications, core-fucosylation and bisecting GlcNAc, were developed based on the GU library. Second, the applicability of this method for the analysis of complex biological samples is evidenced by the ability to discriminate between cell culture and tissue sample types by the normalized intensity of N-glycan structures alone. Overall, the methods and data described here are expected to support the future development of more automated approaches to glycan identification and quantitation. [doi:10.25345/C55G99] [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: glycomics ; N-glycans ; O-glycans

Contact

Principal Investigators:
(in alphabetical order) 		Nicolle H. Packer, Macquarie University, Australia
Submitting User: cashwood

Publications

Ashwood C, Pratt BS, MacLean BX, Gundry RL, Packer NH.
Standardization of PGC-LC-MS-based glycomics for sample specific glycotyping.
Analyst. 2019 May; Just accepted. doi: 10.1039/C9AN00486F.

Christopher Ashwood, Brian Pratt, Brendan MacLean, Rebekah L. Gundry, Nicolle H. Packer.
Standardization of PGC-LC-MS-based glycomics for sample specific glycotyping.
Ashwood, Christopher; Pratt, Brian; MacLean, Brendan; Gundry, Rebekah L.; Packer, Nicolle H. (2019): Standardization of PGC-LC-MS-based glycomics for sample specific glycotyping. ChemRxiv. Preprint. https://doi.org/10.26434/chemrxiv.7056767.v2.

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