Cell-based Glycan Display Array

Uncover the Beauty of Glycan Diversity with Our Cell-based Display Array

CD BioGlyco has a robust Glycan Display Platform that provides clients with a comprehensive range of services related to glycan display arrays. Among them, cell-based glycan display arrays, which are rich in multiple types and diverse contents, form an important part of the glycan display platform.

To make the constructed glycan display arrays stable as well as more widely applicable, we chose the HEK293 cell line, which is easily transfected to express recombinant glycoproteins, to construct glycan display arrays. We generate homozygous cell libraries with different glycosylation abilities through gene knock-in (KI) and knock-out (KO). At the same time, we adopt the " glycan pruning" method to design and optimize the glycosylation capacity of this cell library, and then construct cell-based glycan arrays.

First, we provide combinatorial approaches to eliminate or introduce de novo glycosylation capacity in cell lines to develop a sublibrary of stably engineered HEK293 homozygous cells that individually show loss or gain of different features of the glycomics.
We generate cellular sublibraries for the major steps of glycosylation using an isogenic cell set with the ability to reprogram glycosylation. Among other things, the sublibrary differentially displays major types of glycocouplings by eliminating the pathway-specific extension step of one or more glycoconjugates to display different types of glycans.
We provide targeted KI integration of glycosyltransferase genes to introduce glycan features not endogenously expressed in HEK293 cells. We use KI of dominant glycosyltransferases to enhance the corresponding glycan features.
We use flow cytometry to detect glycosyltransferase genes. Positive or negative signals provided by the loss or gain of KO and/or KI interactions of glycosyltransferase genes are used to interpret the structural features of the glycans and the involved glycoconjugates.
Finally, we employ a range of plant, microbial, and human lectins to demonstrate the performance of the arrays.

Fig.1 Cell-based glycan display array service. (CD BioGlyco)

The specific cell-based glycan display arrays related services we offer but are not limited shown below:

Glyco-engineered Cell Construction

CD BioGlyco employs glycoengineering techniques to efficiently construct cell lines through processes such as the construction of cell sublibraries and the design and regulation of cellular glycosylation capacity. At the same time, we provide glycoconjugate engineering cell construction services according to the different types of glycosylation pathways, including but not limited to the GPI Anchor Pathway, Glycolipids Pathway, N-linked Glycan Pathway, O-GalNAc Pathway, O-Fuc Pathway, O-Man Pathway, C-Man Pathway, O-Glc Pathway, O-Xyl Pathway, O-Gal Pathway, O-GlcNAc Pathway, Glycan Elongaton/branching Pathway, and Glycan Capping Pathway.

Cell-based O-Glycan Array

CD BioGlyco constructs glycan arrays displaying O-glycans based on glycoengineered construct cells. This array displays various types of O-glycans and describes how O-glycans interact with each other.

Cell-based N-Glycan Array

CD BioGlyco utilizes a combinatorial approach to glycobiology to generate an N-glycan array based on the construction of a cellular sublibrary. This array clearly shows the glycan features of N-glycans on the cell surface.

Cell-based Glycosaminoglycan (GAG) Array

CD BioGlyco utilizes a combination of glycoengineering techniques to generate arrays displaying GAGs. This array displays and dissects the biosynthesis as well as the structural basis of GAG.

Cell-based Glycoprotein Array

This is a cell-based glycan array showing homogeneous mucins and glycoproteins produced and purified from cell arrays. Glycoproteins including glycan controls can be used for interaction research.

Publication

Technology: Glycosylation pathway mapping and glycoengineering technique

Journal: Molecular Cell

IF: 16.0

Published: 2019

Results: Glycan arrays display oligosaccharides for reporting glycan hemiantigen binding epitopes. Glycan arrays are a limited resource for the presence of glycans in the absence of other glycans and glycoconjugates. Utilizing glycosylation pathway mapping, the authors generated a library of homozygous HEK293 cells with combinatorially engineered glycosylation ability, which was used to display and dissect the genetic, biological synthesis, and structure basis of glycan binding in the natural environment. With the cell-based glycan array being self-renewable and reporting glycosyltransferase genes required for interactions through logically sequenced biosynthetic steps, the involved structural glycan characteristics are predicted and guide synthesis, recombinant production, and genetically dissected strategies. The broad utility of the cell-based glycan arrays structured by the authors is demonstrated and reveals higher-order incorporation of microbial adhesins into O-glycan tufted plaques that are organized by their presentation on proteins.

Fig.2 Depiction of the KO and/or KI gene engineering strategy for the development of isogenic HEK293 cells with selective loss or gain of glycosylation capacities. (Narimatsu, et al., 2019)

Applications

Cell-based glycan display arrays can be used to investigate and map information about complex glycans.
Cell-based glycan display arrays can be used to probe the glycan binding specificity of lectins and microbial adhesins.
Cell-based glycan display arrays can be used to report complex glycan semi-antigenic structures, to display and interrogate human glycoproteins, and to produce recombinant glycoproteins with desired glycosylation.

Advantages

Our constructed glycan arrays contain multiple types that display customized N- or O-linked glycoforms.
Our constructed cell-based glycan display arrays provide accurate and reliable glycoform prediction and interaction research to ensure client satisfaction.
We have a professional R&D team and every solution we provide is quality tested.

CD BioGlyco provides high-quality glycan display array construction services to clients. Our glycan display arrays can be used to probe the interactions and binding of microorganisms, lectins, antibodies, and other biomolecules to glycans, oligosaccharides, and proteins. If you are interested in our glycan display arrays, please feel free to contact us.

References

Büll, C.; et al. Cell-based glycan arrays-a practical guide to dissect the human glycome. STAR Protocols. 2020, 1(1): 100017.
Narimatsu, Y.; et al. An atlas of human glycosylation pathways enables display of the human glycome by gene engineered cells. Molecular Cell. 2019, 75(20): 394-407.

This service is for Research Use Only, not intended for any clinical use.