Recently isotope labeling and direct quantitative methods have strengthened comparative glycan analysis. CD BioGlyco has been committed to developing various metabolic labeling strategies for many years. We look forward to cooperating with you and becoming your research assistant in glycomics.

Background

Glycan analysis requires high sensitivity and high throughput analytical techniques. Mass spectrometry has been widely used in the study of glycomics, which meets the needs for rapid and reliable information for the analysis of complex glycan mixtures. With the development of quantitative proteomics, several stable isotope labeling strategies in vivo and in vitro have been described for glycan quantitative analysis. Although most in vitro isotopic labeling methods are robust, they usually require additional sample processing steps, resulting in increased experimental errors. Metabolic labeling can circumvent these problems because labeling occurs before any sample treatment.

Isotopic detection of aminosugars with glutamine (IDAWG) is an in vivo metabolic labeling method of glycans. This strategy introduces isotopic nitrogen into all cellular aminosugar-containing glycan structures through hexosamine biosynthetic pathway (HBP). However, due to the limited existence of aminosugars in glycans, it is usually difficult to avoid mass spectrometry interference. To solve this problem, a new relative quantitative method of glycans called metabolic isotope labeling of polysaccharides with isotopic glucose (MILPIG) has emerged, which uses isotopic-carbon containing glucose into HBP to produce isotopic labeled UDP GlcNAc for final glycan qualification.

Fig.1 Strategy of metabolic isotope labeling of polysaccharides with MILPIG. (A) Isotope labeling of glycoconjugates through HBP. (B) Schematic of MILPIG strategy for relative quantification of glycans. (Kim, 2021)

Services

CD BioGlyco has developed several in vivo metabolic labeling strategies for quantitative glycan analysis with high sensitivity. Services we provide include but are not limited to:

• We offer reliable quantification services of glycan expression differences after glycans, which are released from proteins and lipids isolated from differentially labeled samples.
• Quantify N-linked and O-linked glycans by adding isotope-labeled glucose to glucose-free medium. This metabolic labeling strategy can be applied to a variety of cell culture systems.
• Analysis of glycoconjugate fluxes and turnover experiments using pulse-chase approaches.

Applications

• Application in quantifying N-linked and O-linked glycans
• Comparing the global levels of additional glycoconjugates
• Investigating the biological role of glycans in cell culture systems

Advantages of Us

• Accurate qualitative and quantitative results
• High sensitivity based on advanced mass spectrometry platform
• Experienced R&D teams
• Detailed technical guidance

Metabolic labeling is a powerful quantitative tool for studying the biological role of glycans in glycoscience. Several simple and powerful strategies in vivo isotope labeling for quantitative glycan analysis have been developed by CD BioGlyco. We have the confidence to provide high-quality service of glycomic quantitation for our customers. please contact us directly if you have any questions.

References:

1. Kim, J.Y.; et al. Metabolic labeling of glycans with isotopic glucose for quantitative glycomics in yeast. Analytical Biochemistry 2021, 621(3):114152.
2. Orlando, R.; et al. IDAWG: Metabolic incorporation of stable isotope labels for quantitative glycomics of cultured cells. Journal of Proteome Research. 2009, 8(8):3816-3823.