What We Do for N-linked Glycoprotein-based Glycophage Display System Construction Service

At CD BioGlyco, our glycan display platform includes a variety of Glycan Display technologies, including Glycophage Display, etc. The N-linked glycoprotein-based glycophage display system is a method for efficiently enriching phages displaying N-linked glycoproteins. The system allows for the presentation of different glycoprotein structures on the phage surface, allowing for the study of glycan-protein interactions, among others.

We provide a one-stop N-linked glycoprotein-based glycophage display system construction service for the study of bacterial N-glycosylation pathways and other studies. We use Escherichia coli as a host system for the presentation of N-linked glycoproteins on the phage surface by fusing genes encoding glycoproteins and phage shell proteins to each other. Since E. coli lacks a natural glycosylation system, glycophages prepared in E. coli only encode non-glycosylated fusion proteins (N-linked glycoproteins-shell proteins). We construct plasmids encoding genes for relevant enzymes in the N-linked glycosylation pathway, such as functional oligosaccharyltransferase. Then we introduce them into E. coli, resulting in the display of glycosylated fusion proteins on the surface of the phage. This phenotypic display i.e. corresponds to any gene encoding a component of the N-glycan biosynthetic pathway. By this method, the various steps of the N-linked glycosylation pathway are studied. In addition to this, oligosaccharide biosynthesis pathway studies are also realized by the glycophage display system.

Our process of constructing the N-linked glycoprotein-based glycophage display system consists of the following steps:

• Selection of glycoproteins
  We select the appropriate N-linked glycoproteins to be displayed on the phage surface according to client requirements.
• Gene modification
  The gene encoding the glycoprotein is inserted into the phage genome. The glycoprotein gene is usually fused to the phage shell protein gene, thereby allowing the glycoprotein to be displayed on the phage surface.
• Fusion protein expression and phage purification
  During phage maturation, fusion proteins are assembled onto the phage surface. Glycophage formation also requires enzymatic modification of the fusion proteins before they are assembled onto the surface of the phage particle. This modification is achieved by incorporating a plasmid encoding a glycosylase.
  We have developed appropriate high-affinity selection procedures to selectively enrich the resulting progeny phage for glycophage. We increase the percentage of glycophage recoverable by, for example, reducing incomplete glycosylation. During the purification process, we modify the enrichment strategy according to specific needs, e.g. using specific antibodies or lectins to ensure selection of the target structure.
• Validation and characterization
  We use western blotting, immunofluorescence, and enzyme-linked immunosorbent assay (ELISA) techniques to validate and characterize the glycoproteins themselves. The successful construction of the glycophage display system is confirmed by analyzing the N-linked glycoprotein presentation on the phage surface.

Fig.1 Process of N-linked glycoprotein-based glycophage display system construction. (CD BioGlyco)

Publication Data

Applications of N-linked Glycoprotein-based Glycophage Display System Construction

• The N-linked glycoprotein-based glycophage display system is used to identify glycan-binding ligands and to discover therapeutic targets.
• The N-linked glycoprotein-based glycophage display system is used to study the bacterial N-linked glycosylation pathway.
• The N-linked glycoprotein-based glycophage display system is used to gain insight into the structure of specific glycans recognized by different proteins, thus providing a deeper understanding of the role of glycans in various biological processes.

Advantages of Us

• We refine and optimize N-linked glycoprotein-based glycophage display systems according to the needs of our clients. The specific experimental protocols and techniques used may vary.
• We explore different host systems, and phage species, and optimize constructs to enhance the value of the glycophage display system.
• With our rich research and experience in glycophage display, we accomplish various challenges with excellence.

CD BioGlyco has built a team of experts in glycophage display to provide various glycan analysis services. We customize solutions according to client requirements. Please feel free to contact us for more information about the construction of N-linked glycoprotein-based glycophage display system.