Targeted glycomics by selected reaction monitoring for highly sensitive glycan compositional analysis

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DOIResolve DOI: http://doi.org/10.1002/pmic.201100567
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TypeArticle
Journal titleProteomics
ISSN16159853
Volume12
Issue15-16
Pages25102522; # of pages: 13
SubjectCarbohydrate; Glycoproteomics; Glycosylation; LC-SRM; N-Glycan; Quantitative glycomics
AbstractThe development of glycomics increasingly requires the detection and quantification of large numbers of glycans, which is only partially achieved by current glycomics approaches. Taking advantage of selected reaction monitoring to enhance both sensitivity and selectivity, we report here a strategy termed targeted glycomics that enables highly sensitive and consistent identification and quantification of diverse glycans across multiple samples at the same time. In this proof-of-principle study, we validated the method by analyzing global N-glycans expressed in different systems: single proteins, cancer cells, and serum samples. A dynamic range of three orders of magnitude was obtained for the detection of all five glycans released from ribonuclease B. The limit of detection of 80 attomole for Man9GlcNAc2 demonstrated the excellent sensitivity of the method. The capability of the strategy to identify diverse glycans was demonstrated by identification and detection of 162 different glycans and isomers from pancreatic cancer cells. The sensitivity of the method was illustrated further by the ability to detect eight glycans from 250 cancer cells and five glycans released from 100 cancer cells. In serum obtained from rabbits fed control diet or diet enriched with 2% cholesterol, differences to 42 glycans were accurately measured and this indicates that this strategy might find use in studies of biomarker discovery and validation.
Publication date
LanguageEnglish
AffiliationHuman Health Therapeutics; National Research Council Canada
NoteThe authors affiliated with NRC were with the NRC Institute for Biological Sciences prior to April 1, 2012.
Peer reviewedYes
NPARC number21268230
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Record identifieree4d1fa2-6367-4c8f-89c1-d44c07ffeab5
Record created2013-06-06
Record modified2016-05-09
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