Sialic acids are in exposed positions on macromolecules and cell surfaces and are potent modulators of cellular function. Therefore, their turnover or susceptibility to degrading enzymes is of interest for genetic diseases. CD BioGlyco has an efficient Enzymatic Release method to provide clients with sialic acid release service.

Overview of Sialic Acid

Glycosylation is a common post-translational modification that plays a crucial role in protein folding, trafficking, stability, and cellular activity. The two main types of glycosylation are N-glycosylation and O-glycosylation. Sialic acids are usually located on glycan terminal residues of N-glycosylation and O-glycosylation. They are a class of α-keto acid sugars composed of a nine-carbon skeleton and are one of the most important molecules in life. Sialic acids not only play an important role in the chemistry and biodiversity of glycoconjugates but is also closely related to many diseases, such as Salla disease, atherosclerosis, and influenza.

Fig.1 Sialic acid-targeted biointerface materials and bio-applications. (Xiong, et al., 2017)

Sialidase is one of the most important enzymes in sialic acid catabolism. It removes sialic acid residues from the cell surface or serum sialic glycoconjugates. Typically, readily degradable glycoconjugates are captured by endocytosis in higher animals. Lysosomal sialidases remove sialic acid residues when late endosomes fuse with lysosomes. The activity of these sialidases is based on the removal of the O-acetyl group. A special form of sialidase is trans-sialidase, which combines the properties of sialidase and sialyltransferase. This enzyme prefers to transfer a sialic acid molecule from a glycoside to another sugar molecule, forming only an α-2,3 bond.

Since glycoprotein sialylation is critical to the bioavailability, stability, metabolism, and immunogenicity of protein therapeutics. Therefore, sialic acid is often released from glycoproteins in routine protein analysis. The content of sialic acid in the protein is then determined by various spectroscopic and chromatographic methods and its structure is characterized.

Key Technologies

CD BioGlyco leverages a suite of cutting-edge technologies to deliver superior enzymatic sialic acid release services. Our expertise is built upon a foundation of advanced enzymatic tools and state-of-the-art analytical platforms. Key to our success is the careful selection and application of highly specific sialidases tailored to target different glycosidic linkages of sialic acids (e.g., alpha 2,3-, alpha 2,6-, and alpha 2,8-linked sialic acids). This specificity ensures precise removal of sialic acids without affecting the underlying glycan structure.

Beyond enzymatic digestion, our core technologies include:

• High-performance liquid chromatography (HPLC) / ultra-high-performance liquid chromatography (UHPLC)
• Mass spectrometry (MS) / tandem mass spectrometry (MS/MS)
• Capillary electrophoresis (CE)
• Specialized derivatization techniques

Unveiling the Sialic Acid: Precision Enzymatic Release for Unmatched Glycan Insights

Sialic acid determination is performed by a variety of methods. Typically, sialic acid is released from glycoproteins by acid hydrolysis or enzymatic digestion before analysis. At CD BioGlyco, we have developed specialized Glycan Release platforms. To provide clients with efficient and accurate sialic acid release services from native glycoprotein or glycan structures. We completely release α2-3, α2-6, and α2-8-linked sialic acids present on N-glycans and O-glycans under the action of sialidases. In addition, we also provide fast and sensitive Sialic Acid Analysis services through high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD), liquid chromatography-mass spectrometry (LC-MS), etc.

Fig.2 The process of enzymatic release of sialic acid. (CD BioGlyco)

CD BioGlyco's enzymatic release of sialic acid service offers a broad scope, catering to diverse research and industrial needs. Our capabilities extend to:

• Quantitative and qualitative analysis of sialic acids: Determining the exact types and amounts of sialic acids present on various glycoconjugates.
• Sialic acid linkage isomer analysis: Differentiating and quantifying alpha 2,3−, alpha 2,6−, and alpha 2,8-linked sialic acids, critical for understanding their biological functions.
• Analysis of O-acetylated sialic acids: While challenging due to their lability, our optimized methods allow for the detection and characterization of O-acetylated sialic acids, which are increasingly recognized for their roles in health and disease.

Workflow

Publication Data

Applications

• Biopharmaceutical characterization: Critical for the quality control and characterization of glycoprotein therapeutics (e.g., antibodies, recombinant proteins), where sialylation profoundly impacts efficacy, pharmacokinetics, and immunogenicity.
• Biomarker discovery and diagnostics: Analyzing changes in sialylation patterns in various biological fluids or tissues reveals potential biomarkers for diseases such as cancer, inflammatory disorders, and infectious diseases.
• Glycan structure elucidation: As a crucial preparatory step, enzymatic release allows for the subsequent detailed structural analysis of released sialic acids and the underlying desialylated glycans, contributing to a comprehensive understanding of glycan architecture.

Advantages

• Unrivaled specificity and purity: Unlike harsh chemical methods that lead to non-specific cleavage or degradation of glycans, our enzymatic approach ensures highly specific release of sialic acids, minimizing undesirable side reactions.
• Preservation of glycan integrity: The mild, controlled conditions of enzymatic digestion safeguard the underlying glycan structures, enabling subsequent comprehensive glycan analysis without concerns about structural alterations.
• Comprehensive linkage specificity: CD BioGlyco offers an extensive panel of highly characterized sialidases, allowing for targeted release of specific sialic acid linkages (alpha2-3, alpha2-6, alpha2-8,etc.).

Frequently Asked Questions

At CD BioGlyco, we are dedicated to advancing glycobiology research through our specialized services and cutting-edge technologies. Our enzymatic release of sialic acid service provides a precise, reliable, and gentle method for dissecting the complex world of sialylation, offering invaluable data for biopharmaceutical characterization, biomarker discovery, and fundamental biological research. Contact us for more information and to discuss your project.

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