Cell-based O-Glycan Array

Cell-based O-Glycan Array

Precision and High Throughput for Your Cell-based O-Glycan Array

CD BioGlyco is dedicated to helping clients explore the development of glycan biological functions. As a leading biotechnology company in the field of glycomics, we comprehensively dissect molecular interactions and biological functions of glycans through a wide range of biological analysis services. The cell-based O-glycan array is composed of a series of Glyco-engineered Cell lines (HEK293 cells), which express homogeneous and personalized O-glycans in cellular expression and are used for validation of conformational relationships.

Cell surface O-glycans have structural diversity and complexity. The binding of cell surface O-glycosylation capacity with ligands provides a means for studying the molecular recognition mechanism. Meanwhile, CD BioGlyco provides more comprehensive and systematic engineering development services based on cell-based O-glycan arrays displaying glycocell bio-interrogation. We provide services ranging from cell-based O-glycan array construction to biosynthesis genetic regulation metabolic analysis.

  • Construction service

High-purity O-glycans are key to building arrays. At the glycan display platform, we provide a series of HEK293 cells genetically engineered to express O-glycans with specific structures e.g. O-GalNAc, O-Fuc, O-Man, O-Glc, O-Xyl, O-Gal and O-GlcNAc. Our researchers have a richer experience with array expansion to ensure fast array construction and validation.

  • Selecting and designing service

Our researchers provide fast and accurate selection services to help clients pick out glycan-binding proteins (GBP) or glycan-binding reagents. Based on the information (known / partially known / unknown), we provide corresponding services. Our talent team has a rich background in glycobiology and rich experience in industry services to grasp every key node.

  • Known glycan epitope: we validate the binding properties by using a glycosylation signature library. Moreover, we provide analysis services for glycan epitope isogenic cell interaction.
  • Partially known glycan epitope: we provide a detailed analysis service by selecting a gene-editing library containing (non-) specific glycosyltransferase (GTf) genes for pathways associated with glycan epitope.
  • Unknown glycan epitope: we perform binding validation services by testing its binding properties to the HEK293 cell line and evaluating the major types of glycoconjugate libraries. Notably, if it is not bound to the HEK293 cell line or bound to other cell lines, we provide the GTf gene expression detection service.
  • High-throughput analysis service

Combined with a large number of isogenic cells and cell-based O-glycan arrays, we provide binding-specific detection services by high-throughput flow cytometry. We provide surface binding, membrane localization, density, or aggregation assessment services by immunohistochemistry. Furthermore, we provide live cells to immobilized O-glycan recognition services.

Fig.1 Schematic diagram of cell-based O-glycan array analysis (CD BioGlyco)Fig.1 Schematic diagram of cell-based O-glycan array screen and analysis (CD BioGlyco)

Publication

Paper Title: Installation of O-glycan sulfation capacities in human HEK293 cells for display of sulfated mucins

Technology: RNA sequencing, Knockin, Knockout, Flow cytometry

Journal: Journal of Biological Chemistry

Published: 2022

IF: 5.485

Results: In this paper, a cell-based glycan array was extended by a sialylation-competent HEK293 cell line. Endogenous glycan branching and sialylation capacity were eliminated by targeted knockout of the sulfotransferase gene in combination with knockout genes to provide a simplified substrate introduction of sulfotransferases. In addition, analysis of lectin binding specificity using engineered cell libraries indicated binding to the 3-O-sulfo-core1 O-glycan. Importantly, a cell-based platform was used to display and produce glycoconjugates of sulfated O-glycans. This strategy is generalizable to other sulfotransferases.

Fig.2 Intact mass analysis of the Lys-C-purified mucin1 TR O-glycodomain. (Sun, et al., 2022)Fig.2 Intact mass analysis of the Lys-C-purified mucin1 TR O-glycodomain. (Sun, et al., 2022)

Applications of Cell-based O-Glycan Array

  • Cell-based O-glycan array can be used to discover functional glycans, characterize glycan processing enzymes, and detect pathogens for diagnosis.
  • Cell-based O-glycan array provides a broader range of glycan epitopes and allows the recognition of native ligands.
  • Cell-based O-glycan array advances the specific development of GTf genetically engineering linker libraries for in-depth analysis of reagent binding.
  • Cell-based O-glycan array is suitable for monitoring the interaction of multiple molecules with biomolecules.
  • Cell-based O-glycan array effectively recognizes structural sugar features recognized by GBP or other glycan-binding reagents and by the GTf gene.

Advantages of Us

  • Our investigators use fixed cells instead of live cells for flow cytometry analysis, avoiding extensive cell culture and processing. We optimize the service process and speed up the project's progress.
  • Our operator performs staining services at 4°C to avoid the uptake of reagents or fluorophores and to avoid some lectin-induced apoptotic signals due to glycan crosslinking.

CD BioGlyco has a professional Glycan Display Platform to provide clients with Cell-based Glycan Display Array services. We are committed to being your best partner in the field of glycobiology. To learn more information about cell-based O-glycan arrays, please feel free to contact us.

References

  1. Nason, R.; et al. Display of the human mucinome with defined O-glycans by gene engineered cells. Nat Commun. 2021, 12(1): 4070.
  2. Sun, L.; et al. Installation of O-glycan sulfation capacities in human HEK293 cells for display of sulfated mucins. J Biol Chem. 2022, 298(2): 101382.
This service is for Research Use Only, not intended for any clinical use.

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CD BioGlyco is a world-class biotechnology company with offices in many countries. Our products and services provide a viable option to what is otherwise available.

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