Sialyltransferases (SATs)-based Cell Surface Chemoenzymatic Glycoengineering Services at CD BioGlyco

SATs are enzymes that catalyze the transfer of sialic acid residues to glycan structures, which play a vital role in cell surface glycoengineering, and CD BioGlyco provides mature SATs-based cell surface chemoenzymatic glycoengineering service.

• Enzyme production and purification services

CD BioGlyco provides high-quality SAT production and purification services. SATs are key enzyme that catalyzes the transfer of sialic acid residues from cytidine 5'-monophosphate-sialic acid (CMP-sialic acid) to acceptors, which is commonly applied in the synthesis of sialic acid-containing structures. For example, a highly specific α2-3-sialyltransferase named PmST1_M144D from Pasteurella multocida generates the sialic acid Lewis X structure. We use molecular chromatography techniques, such as affinity chromatography, gel filtration chromatography, etc., to achieve further purification of PmST1_M144D based on the properties and affinity.

• Cell chemical enzyme labeling services

CD BioGlyco incubates PmST1_M144D with the substrate CMP-Neu5Az in tris buffer, using several cell surface glycans of Chinese hamster ovary (CHO) cells as receptors. PmST1_M144D binds Neu5Az to surface glycoconjugates with N-acetyllactosamine (LacNAc)-terminal substrates in cells, enabling efficient sialylation. At the same time, we provide a variety of different types of sialic acid modifications introduced based on SAT, such as N-acetyl sialic acid (Neu5Ac) and N-methyl sialic acid (Neu5Gc), thus expanding the diversity and functionality of glycosyl structures.

• Glycosylation analysis and characterization services

CD BioGlyco provides glycosyl analysis and characterization of cell samples that have been treated with chemical enzymatic glycoengineering. Including mass spectrometry, lectin binding experiments, flow cytometry, and other techniques to evaluate the effects and changes of glycosyl modifications. We also use specific markers or fluorescent probes to label the introduced sialic acid modifications to enable detection, tracking, and quantitative analysis of the modifications.

Fig.1 SATs-based cell surface chemoenzymatic glycoengineering services. (CD BioGlyco)

Publication

Technology: Chemoenzymatic glycoengineering approach

Journal: Biology Open

IF: 2.643

Published: 2017

Results: The authors used a highly specific sialyltransferase (PmST1_M144D) to label living CHO cells. The results showed that PmST1_M144D could effectively sialylate LacNAc, and CHO cells achieved efficient glycosylation, which was verified by bioorthogonal blotting experiments.

Fig.2 CHO cells labeled by chemoenzymatic glycoengineering. (Almaraz & Li, 2017)

Applications

• Cell biology research: CGE facilitates the study of cell biology by providing an alternative method to specifically label glycans on the surface of living cells, enhancing the ability to visualize the glycome, which will greatly aid the study of glycoprotein function in living systems.
• Vaccine research: SATs introduce specific sialic acid modifications on the cell surface, thereby changing the cell surface sugar structure. This glycoengineering strategy is applied to develop sialic acid-modified vaccines to improve vaccine immunogenicity and efficacy.
• Study on glycosyl function: SATs introduce different types of sialic acid modifications on the cell surface to simulate the diversity of glycosyl groups in nature, which helps to study the effects of different glycosyl structures on cellular function, molecular recognition, and interactions, thereby revealing the biological functions and mechanisms of glycosyls.

Advantages

• SATs are applied to a variety of cell types, CD BioGlyco provides different species of cell surface chemoenzymatic glycoengineering services, including common mammalian cells and microorganisms.
• CD BioGlyco provides efficient cell surface chemoenzymatic glycoengineering services. We use SATs to quickly introduce targeted glycosyl modifications in a relatively short period of time.
• CD BioGlyco performs personalized chemoenzymatic sugar engineering design for different types of cells and substrates, which has the advantages of diversity and flexibility.

CD BioGlyco has a mature Cell Surface Glycoengineering technology platform, and we provide professional SAT-based cell surface chemoenzymatic glycoengineering services. Please feel free to contact us in time if you would like to consult the relevant service details.