Acetylated Sialic Acids-based Cell Surface Glycoengineering Service at CD BioGlyco

CD BioGlyco has a well-established Platform for Glycoengineering. Sialic acid molecules are rich in active hydroxyl groups, which are often replaced by O-acetyl groups. O-acetylation is one of the common modifications of sialic acid. This structure is frequently found on glycoproteins on the surface of cell membranes and glycoproteins secreted outside the cell, especially at the ends of the sugar chains of N- and O-sugars. In order to meet the needs of different studies targeting acetylated sialic acid, we have developed a Cell Surface Glycoengineering service based on acetylated sialic acid. Current studies on the biological functions of the O-acetylation modification of sialic acid are focused on cancer development and progression, embryonic development and immune processes, drug targeting, etc. The glycoengineering services we offer will meet the needs of our clients for different studies.

Fig.1 Types of our cell surface glycoengineering service based on acetylated sialic acid. (CD BioGlyco)

• Targeted introduction or modification of acetylated sialic acid

We alter the structure of acetylated sialic acid by chemical modification. We preferentially O-acetylate on the hydroxyl groups of salivary acid carbon atoms C4, C7, C8 and C9. In addition to this, we regulate the synthesis of acetylated sialic acid by Gene Editing, thus targeting the introduction of acetylated sialic acid with a specific structure. Our goal is to enrich the diversity of salivary acids on the cell surface by adding multiple types of acetylated sialic acid.

• Quantitative analysis service for acetylated sialic acid

One of the most widely used methods for the quantitative Analysis of Salivary Acids is the combination of liquid chromatography and mass spectrometry. We have vast experience in the quantitative analysis of salivary acids. Salivary acids are located at the end of sugar chains such as glycoproteins. They need to be released before analysis. Salivary acid release involves two methods: acid hydrolysis and enzymatic hydrolysis. We release salivary acid from samples by acid hydrolysis.

Currently, derivatization based on 1,2-diamino-4,5-methylenedioxybenzene (DMB) is the routine method for the quantification of biological samples and glycoproteins with salivary acid. The benefits of this derivatization are twofold. First, it attaches fluorescent groups to the sialic acid, thereby facilitating its analysis by liquid chromatography-fluorescence detector. Second, it enhances the ionization efficiency of salivary acid during mass spectrometry analysis, thereby enhancing its detection sensitivity. We employ DMB for fluorescent labeling of sialic acid. After the derivatization reaction, the derivative is detected and analyzed by reversed-phase (U) high-performance liquid chromatography coupled with mass spectrometry.

Publication

Technology: Liquid Chromatography–Orbitrap Mass Spectrometry (LC–Orbitrap-MS)

Journal: Analytical Chemistry

IF: 6.785

Published: 2019

Results: In this study, a method for the analysis of acetylated sialic acid based on dried blood spot sampling (DBS) was developed using crucian carp blood as experimental material. Sialic acid was first released from DBS by acid hydrolysis. Then the released sialic acid was fluorescently labeled with DMB. Finally, the labeled salivary acids were analyzed by LC-Orbitrap-MS. The LC-MS results showed that the salivary acids in crucian carp blood were structurally diverse. Among them, O-acetylated substances were detected as multiple peaks. The results suggest that salivary acid heterogeneity is caused by a combination of the number of O-acetyl groups and their positions.

Fig.2 LC-MS analysis of DMB-derivatized sialic acids from crucian carp blood. (Zhang, et al., 2019)

Applications of Our Acetylated Sialic Acids-based Cell Surface Glycoengineering Service

• Acetylated salivary acid has important functions in embryogenesis, development, and other processes. Cell surface glycoengineering based on acetylated salivary acids helps to study the role of salivary acids in life activities.
• Cell surface glycoengineering based on acetylated sialic acid can be used to analyze cell surface acetylation.
• Acetylated salivary acids may be potential disease markers. Cell surface glycoengineering based on acetylated sialic acid can be used to analyze the mechanisms of disease development such as cancer.

Highlights of Us

• The glycoengineering methods we construct for acetylated sialic acid have a wide range of applications. Acetylated salivary acids can be analyzed in simple or complex biological samples.
• Detection and analysis of O-acetylated sialic acid requires multiple precautions in sample preparation. Care is taken to avoid loss of O-acetyl groups during experiments to ensure accurate results.

CD BioGlyco is experienced in the glycoengineering of salivary acids. Neu5Gc, Neu5Ac, and acetylated salivary acids can be researched. We aim to facilitate our clients in glycobiology research. No matter what the aspect, our goal is always to provide the best quality service. Please feel free to contact us to ask for more information about glycoengineering of salivary acids and more.