Empowering Glycans for Innovative Discoveries

Cell construction is a critical step in the development of Cell-based Glyco-display Arrays, and CD BioGlyco has best-in-class glycoengineering techniques to provide trusted cell construction services.

• Primary cell banks: To expand the scope of the cells for subsequent applications, we use human embryonic kidney 293 (HEK293) cells, a cell line widely used for glycoproteins, as the basis for cell construction.
• Working cell banks: We perform a culture of isogenic HEK293 cells. We culture HEK293 cells in adherent wall culture or suspension culture to generate isogenic cells, i.e., the master cell bank. In addition, we construct isogenic cells with specific glycosylation features by altering the glycan complexity branching through the knockout and knock-in of glycosyltransferase genes. Among them, adherent HEK293 cells are mainly used for binding research with or without transient expression of membrane glycoprotein reporter genes, while the mixed suspension HEK293 is more favorable for high-yield production of recombinant glycoproteins.

Alternatively, glycoprotein reporter genes can be designed for transient expression in homozygous cells. We transiently transfected glycoprotein reporter genes into homozygous HEK293 cells using a well plate format. We confirm the expression of the transfected glycoproteins by co-expression of fluorescent reporter proteins encoded in the expression plasmids or other techniques (e.g. enzyme-linked immunosorbent assay, sodium dodecyl sulfate-polyacrylamide gel electrophoresis).

Next, we use live cells for flow cytometry analysis. We utilize the dispensing of vials or 96-well plates with sub-libraries to facilitate analysis in cell construction experiments.

• In addition, we offer an analysis of cell banks:
  We use RNA sequencing (RNA-seq) transcriptomics to predict glycosylation capacity, pathway-specific extension, and branching characteristics of constructed cell libraries. Glycosyltransferase-based CRISPR / Cas9 gene editing tools are provided to design and analyze the glycosylation capacity of cell lines. Use precision genetic engineering to profile and reprogram glycosylation pathways in cell lines. Utilize syngeneic cell models to assess glycosylation pathway mapping.

Fig.1 Glyco-engineered cell construction service. (CD BioGlyco)

Depending on the glycosylation pathway, we offer glycoengineered cell construction services that include, but are not limited to:

Publication

Applications

• Glyco-engineered cell construct can be used to explore glycan and protein interactomes.
• A major step in the creation of a glycan microarray from cells is cell banking. Therefore, glyco-engineered cell construct can be used to produce a synthesis of glycans, predict the structural features associated with their formation, and guide methods for making microarrays, recombinant production, or genetic dissection.
• Glyco-engineered cell construct can be used to probe the possible binding of microbial adhesins to clustered patches of O-glycans.

Advantages

• We have a powerful Glycan Display Platform to provide clients with full-process services such as the construction of cell lines and glycan microarray development services.
• We have mature technical tools to support the research of the interaction between glycans and cells, high-throughput screening, and other processes.
• We analyze and evaluate the constructed cell libraries in all aspects to ensure the quality and validity of the cell libraries.

CD BioGlyco has an experienced R&D team skilled in glycoengineering techniques for cell construction. Each of our protocols is developed by experts in the field of glycobiology to ensure that we provide our clients with high-quality, stable, and safe cell banks. Please feel free to contact us if you would like to inquire about cell construction services.