What Is High Mannose N-glycan?

Glycans on the surface of organisms are important mediators of cell adhesion, signaling, and bacterial-host interactions. N-linked high-mannose glycans not only act as biomass or glycan shields on viral particles (e.g., SARS-COV-2 and HIV) to evade host immune surveillance but also be closely associated with tumor progression when expressed at elevated levels on the surface of cancer cells. Therefore, high mannose-type N-glycans are characteristic structures for immune evasion in many viruses and cancer cells and become potential targets for viral suppression and cancer diagnosis or therapy. On the other hand, high-mannose type N-glycans also have gastrointestinal protection and immunity-enhancing effects.

Cell Surface High-mannose Type N-glycan Glycoengineering Service at CD BioGlyco

CD BioGlyco has state-of-the-art technology for glycan engineering research, and we offer comprehensive services in all aspects of structure, distribution, construction, expression, and application of high-mannose type N-glycans.

• The N-glycan chains of high-mannose type N-glycans contain only a-mannose residues in addition to the core pentasaccharide, with two pentasaccharide structures often being their common core. To analyze site-specific protein glycosylation, we offer mass spectrometry, lectin affinity chromatography (LAC), and hydrophilic interaction chromatography for complete N-glycan chains. In addition, our MALDI-TOF technology has a high sensitivity for the structural characterization of glycosylated compounds and enables high-throughput detection.
• In eukaryotic cells, N-glycosylation (N-Glycan) is carried out in two specialized organelles, the endoplasmic reticulum (ER) and the Golgi apparatus. At CD BioGlyco, we provide services in the study of N-glycosylation (N-Glycan) in living organisms, which leads to the determination of the type of N-glycans on the cell surface.
• CD BioGlyco also offers glycomics reagents. We release most high mannose-type N-glycans from their protein scaffolds. This includes isolating the N-glycan from the tissue, purifying it, separating it on a column, printing it onto microarrays or beads, and then interrogating it.
• We also offer to alter the number and position of glycosylation sites of high mannose-type N-glycans to achieve a change in the structure of the glycan. We also offer N-glycan-based glycoengineering to construct various glycoproteins.

Fig.1 Analytical content of N-glycans. (CD BioGlyco)

Publication

Paper Title: Glycoengineering of HEK293 cells to produce high-mannose type N-glycan structures.

Technology: Expression of recombinant LIPA in KO cell lines

Journal: The Journal of Biochemistry

Published: 2019

Results: To investigate N-glycan structures of recombinant proteins produced in the constructed QD-KO and QT-KO cell lines, human LIPA tagged with a His-FLAG (HFLIPA) was transiently expressed in these cell lines and purified from the medium by anti-FLAG M2 affinity gel. HEK293 WT and T-KO cell lines were subjected to the same experiment as controls. LIPA proteins produced in T-KO, QD-KO, QT-KO, and MGAT1-KO cell lines were detected at 60 kDa, while both EndoH- and PNGaseF-treated LIPA migrated at a lower molecular weight, indicating that the high-Man-type N-glycans became the major glycan form of LIPA produced in these cell lines.

Applications

• Glycoengineering techniques for N-glycans can be used to determine glycan composition and peptide sequences at glycosylation sites.
• Analyzing specific glycan species or determining the relative amounts of specific groups of glycans can be applied to the development of biotherapeutic drugs.
• The monosaccharide composition of glycans can be applied to determine the number and composition of sugar units bound to proteins.

Advantages

• We have a complete glycoengineering system.
• We provide detailed N-glycan glycoengineering services to our clients.
• Our researchers have extensive experience in glycobiology.

CD BioGlyco has a comprehensive, efficient, and stable glycoengineering system for glycan expression, editing, recombination, and analysis. If you are interested in our glycoengineering services, please feel free to contact us.