Specialized Glycoconjugate Analysis Services

Introduction to Glycosylation in Biomolecules

Glycosylation is a crucial post-translational modification impacting the structure, function, and biological activity of various biomolecules, including proteins, lipids, peptides, nucleic acids and small molecules like flavonoids and cannabinoids. For instance:

• Glycosylated flavonoids (e.g., quercetin glycosides) exhibit improved water solubility and bioavailability compared to their aglycone forms, making them promising candidates for anti-inflammatory and neuroprotective therapies.
• Glycosylated cannabinoids, such as cannabosides generated via enzymatic glycosylation, overcome the hydrophobicity of parent cannabinoids, enabling novel oral drug delivery strategies.

Fig.1 Beneficial glycosylated plant metabolites for humans.¹

This process creates complex glycoconjugates like glycoproteins, glycolipids, and glycopeptides. Analyzing these molecules is challenging due to their diverse sugar compositions, linkage types, and structural arrangements. Accurately characterizing glycosylation requires specialized expertise to understand structural variations, identify modification sites, and quantify interactions between glycans and proteins. At Creative Biolabs, we offer specialized glycoconjugate analysis services designed to tackle these complexities. We recognize that each type of biomolecule needs a specific approach to fully understand its glycosylation patterns. Whether you're working with N-glycosylated therapeutic proteins, O-glycosylated cell surface receptors, or glycolipids involved in cell signaling, our comprehensive services utilize advanced technologies to provide precise, useful data. By combining our expertise in molecular diversity with analytical excellence, we empower researchers and biotech partners to navigate the complex field of glycoconjugates, accelerating breakthroughs in glycobiology, disease research, and the development of therapeutics.

Comprehensive Glycosylation Analysis Methods

Glycosylation is a complex post-translational modification that requires careful, specialized techniques to analyze. The variety of glycan structures and glycoprotein differences make this challenging. To understand glycosylation fully, we use a mix of methods. Each method offers unique insights into different aspects, such as glycan composition or modification sites. At Creative Biolabs, our specialized glycoconjugate analysis services combine these techniques to deliver accurate, detailed results for your research needs.

Mass Spectrometry (MS)-Based Methods

MS is a key tool for glycosylation analysis. It offers high sensitivity and accuracy, essential for studying complex glycans. These methods provide both qualitative and quantitative data, helping identify glycan types, modification sites, and protein interactions.

• HILIC-MS is ideal for analyzing polar glycans in glycosylated flavonoids, such as the sialylated or fucosylated derivatives of quercetin. This method, combined with our high-throughput screening platforms, enables rapid profiling of complex samples like plant extracts or microbial fermentation broths.
• MALDI-MS and ESI-MS are used to characterize intact glycoproteins and glycopeptides. For example, alpha glycosyl isoquercitrin, a glycosylated flavonol, can be analyzed via ESI-MS to determine its glycan composition and linkage patterns.

Enzymatic Techniques

Enzymes play a key role in releasing and modifying glycans for analysis. They allow specific cleavage of glycosylation sites, separating glycans from protein backbones.

PNGase F Digestion

• Specifically removes N-linked glycans from asparagine residues in glycoproteins.
• Enables free glycan analysis, especially useful for complex structures with sialic acid or fucose.

O-Glycan Release

• O-glycans attach to serine or threonine residues and are harder to release due to enzymatic resistance.
• Uses O-glycosidases or chemical treatments (e.g., reductive β-elimination) to cleave and isolate O-glycans.
• Follow-up MS or HPLC analysis determines their composition and structure.

Glycobiology Microarrays

Lectin arrays are used to study glycan-protein interactions in glycosylated quercetin and alpha glycosyl isoquercitrin, uncovering their binding specificities to immune receptors or microbial lectins. This is invaluable for optimizing drug-targeting strategies. The high-throughput tool study glycan-protein interactions by immobilizing glycans on a solid surface. Our glycobiology microarray services help uncover glycan-binding specificities for the development of novel biomarkers or therapeutics.

• Immobilize lectins (glycan-binding proteins) to identify specific glycan structures they recognize.
• Screen glycan profiles in samples like serum or cell surfaces, valuable for cancer, immune, and pathogen research.

High-Throughput Screening (HTS) for Glycan

Automated systems process hundreds of samples quickly, integrating techniques like HILIC-MS and glycan microarrays.

• Ideal for large-scale datasets to profile glycosylation patterns efficiently.
• Provides comprehensive results in less time than traditional methods.

Structural Glycomics and Computational Tools

Advanced computational tools and databases predict glycosylation sites and structures using molecular modeling. We use complement experimental methods to offer a complete view of glycosylation's role in cells and disease. Our team combines computational predictions with wet-lab analysis for robust, multi-faceted glycoconjugate characterization.

Glycosylation and Disease: Understanding the Connections

Glycosylation, the enzymatic attachment of sugar molecules to proteins, lipids, and other biomolecules, is a fundamental process in cellular biology. Aberrant glycosylation patterns have been linked to diverse pathological conditions, with specific glycoconjugates playing critical roles in disease progression, diagnosis, and therapy. Below is a disease-specific analysis of glycoconjugates.

Why Choose Us?

• Our experienced team specializes in complex glycosylation analysis of proteins, antibodies, lipids, and small molecules.
• We use the latest mass spectrometry, glycan arrays, and enzymatic tools for precise glycosylation profiling.
• Our services are tailored to meet the unique needs of your research, whether for basic or biopharmaceutical studies.
• We offer high-throughput analysis, ensuring fast processing and efficient research for large-scale studies and drug development.

Glycosylation plays a crucial role in a wide variety of biological processes and is integral to the development and efficacy of many biopharmaceuticals. Our specialized glycoconjugate analysis service offers the expertise and advanced tools necessary to navigate the complexities of glycosylation, ensuring that your research is built on the most reliable data available. Whether you're studying disease mechanisms, developing new therapeutics, or optimizing glycoprotein-based drugs, Creative Biolabs is your trusted partner in glycoscience.

References

1. Kytidou, Kassiani, et al. "Plant glycosides and glycosidases: a treasure-trove for therapeutics." Frontiers in plant science 11 (2020): 357. Distributed under Open Access license CC BY 4.0, without modification. https://doi.org/10.3389/fpls.2020.00357
2. Khodzhaieva, Ruslana S., et al. "Progress and achievements in glycosylation of flavonoids." Frontiers in Chemistry 9 (2021): 637994. https://doi.org/10.3389/fchem.2021.637994
3. Sordon, Sandra, Jarosław Popłoński, and E. W. A. Huszcza. "Microbial glycosylation of flavonoids." Polish journal of microbiology 65.2 (2016): 7. https://doi.org/10.5604/17331331.1204473

Resources

• N-Glycosylation Analysis
• O-Glycosylation Analysis
• DNA Glycosylase and GlycoRNA Analysis
• Fc Region Glycosylation Analysis
• High-Throughput Glycomic Analysis
• Glycosylation Inhibition

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