N-Acetylgalactosamine (GalNAc) release is a key segment in glycomics research. With proven enzyme release methods and comprehensive analytical tools, CD BioGlyco provides clients with high-quality GalNAc release services. We are looking forward to being your experimental assistant in the field of enzymatic release.

GalNAc

Most human-secreted proteins and cell surface proteins are modified by O-GalNAc glycosylation. GalNAc is often the first monosaccharide linked to serine or threonine (Ser/Thr) in a specific form of protein O-glycosylation. O-GalNAc glycosylation is involved in almost every aspect of biology, such as intercellular communication, adhesion, signal transduction, immune surveillance, etc. O-GalNAc glycans are also concentrated in the sensory neural structures of animals and humans and are also terminal carbohydrates that form human blood group antigens. Furthermore, O-GalNAc glycosylation is of fundamental importance in health and disease.

It has been reported that N-acetylglucosamine (GlcNAc) is an inhibitor of elastase released by human polymorphonuclear leukocytes, with inhibition ranging from 8% to 17%. The inhibition range of GalNAc is from 92% to 100%. Therefore, GalNAc is used as a targeting ligand to study liver-targeted antisense oligonucleotides and siRNAs for therapeutic purposes. Aberrant O-GalNAc glycans are a well-established feature of cancer cells and cancer-associated O-glycans, representing a major target for cancer diagnosis and therapy. In conclusion, uncovering the well-defined functions of O-glycans at individual sites in proteins helps better develop site-specific glycan-targeted therapies.

Key Technologies

• Size-exclusion chromatography (SEC): Used as a purification step or for a preliminary separation of glycans by their hydrodynamic volume.
• High-performance liquid chromatography (HPLC): Separates the glycan mixture based on various properties depending on the column chemistry used.
• Mass spectrometry (MS): Provides precise information about the molecular weight and composition of each glycan.
• Capillary electrophoresis (CE): Separates glycans based on their charge and size-to-charge ratio.

Unlocking Glycan Insights: Precision GalNAc Release for Advanced Research

At CD BioGlyco, we utilize a suite of advanced core technologies and methods to ensure high specificity, efficiency, and reproducibility in the enzymatic digestion of GalNAc. Our services include:

• Sample Preparation

Client's samples undergo preliminary purification to remove contaminants that may interfere with enzyme activity or downstream analysis. Sample integrity, concentration, and suitability for enzymatic treatment are also assessed to ensure optimal starting material for digestion.

• Enzyme Selection and Reaction Optimization

We utilize a library of highly specific enzymes, including various α-N-acetylgalactosaminidase and β-N-acetylgalactosaminidase enzymes, each optimized for different linkage specificities and reaction conditions. We also employ enzyme engineering techniques to develop or engineer enzymes with high specificity and activity.

• Enzymatic Digestion

A specific enzyme is incubated with the prepared sample under optimized conditions. The enzyme specifically cleaves the glycosidic bond connecting the GalNAc residue to the sugar chain, releasing the free GalNAc. Our controlled environment ensures consistent and reproducible enzyme activity.

• Product Separation and Purification

After enzymatic digestion, the released GalNAc is separated from the remaining deglycosylated or modified glycans. Techniques such as SPE or SEC are used to separate the digested target component from the enzyme and other reaction byproducts, ensuring a clean sample for subsequent analysis.

• Structural Characterization and Quantification

Advanced analytical methods such as HPLC, MS, and CE are used to comprehensively analyze the released GalNAc and modified glycan structures to confirm the successful and specific release of the GalNAc.

Workflow

Applications

• This service can be used for glycoprotein and glycolipid characterization, which is crucial for elucidating the detailed structure, composition, and linkage types of O-linked glycans and glycolipids.
• This service can be used to identify specific GalNAc-containing glycans that could serve as disease biomarkers, particularly in cancer and inflammatory diseases.
• This service is crucial for characterizing therapeutic glycoproteins (e.g., antibodies and fusion proteins) to ensure consistent glycosylation patterns and assess batch-to-batch variability.
• This service facilitates the analysis of pathogen glycan antigen structures to design effective glycan vaccines.

Advantages

• Unrivaled specificity and precision: Our use of highly purified and linkage-specific N-acetylgalactosaminidases ensures that only the target GalNAc residues are cleaved, minimizing non-specific reactions and preserving the integrity of the remaining glycan structures.
• Gentle and non-destructive approach: Unlike harsh chemical methods that lead to glycan degradation or structural alterations, enzymatic release is performed under mild conditions. This gentle approach maintains the native conformation of the glycans and proteins, providing more biologically relevant data.
• High efficiency and yield: Through extensive optimization of reaction parameters, CD BioGlyco achieves exceptional efficiency in GalNAc release, leading to high yields of the released monosaccharide and modified glycans.

Frequently Asked Questions

CD BioGlyco stands as your premier partner in advanced glycobiology research, offering unparalleled expertise in the enzymatic release of GalNAc. Ready to advance your glycobiology research? Our team of specialists is eager to discuss your specific project requirements and provide a tailored solution. Please feel free to contact us.

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