Innovative O-linked Glycoprotein Display by Glycophages

At CD BioGlyco, our goal is to characterize structurally diverse glycans and glycoconjugates through a variety of Glycan Display techniques. Glycophage Display systems provide an effective tool for the study of O-linked glycoproteins. We provide a glycophage display system construction service based on O-linked glycoproteins. The constructed glycophage display system is used for genetic analysis of O-linked glycoproteins and so on.

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

The establishment of this system involves the construction of glycosylation pathways to enable the presentation of O-linked glycoproteins in phage particles. We combine molecular biology, chemical methods, and glycosylation engineering to purposefully and rationally construct the glycosylation system. The lack of a natural glycosylation pathway in Escherichia coli provides a natural canvas for the construction of O-linked glycosylation. We assemble the formulated glycosylation pathway on it and did not have to worry about interference from endogenous glycosylases. Combined with its ease of genetic manipulation, rapid growth, and ability to express a wide range of recombinant proteins, we use E. coli to produce and clone modified phages. It is found that some bacteria in Pseudomonas and Neisseria have O-linked protein glycosylation pathways. We attempt to form glycosylated fusion proteins by introducing a plasmid encoding a glycosylase for O-linked glycoproteins during phage transformation.

Our construction process includes the following steps:

• Phage construction
  Phage particles are packaged using molecular genetics to express non-glycosylated fusion proteins (phage shell proteins -glycoproteins to be displayed fusion formation). All phages are identified by DNA sequencing. Plasmids encoding O-linked glycoprotein-associated glycosylases are also constructed.
• Production of phages displaying glycosylated glycoproteins
  E. coli monoclonal colonies carrying phages expressing fusion proteins and plasmids encoding glycosylation enzymes are inoculated into the medium and cultured.
• Enrichment of glycophages
  Sugar phages are enriched by lectin affinity chromatography and non-glycosylated sugar phages are screened and removed.
• Western blot analysis
  We characterize glycoproteins on the surface of glycophages by western blotting.

Publication Data

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

• Glycophage display system constructs based on O-linked glycoproteins are used to study the glycosylation pathway.
• O-linked glycoprotein-based glycophage display system constructs are used to establish a physical link between glycoproteins and genes.
• O-linked glycoproteins-based glycophage display system constructs are used to study the information exchange of glycoproteins between cells and their external environment.

Advantages of Us

• We provide an effective tool for high-throughput studies of O-linked glycoprotein interactions by optimizing glycophage display arrays.
• Specific experiments and optimization protocols for the construction of O-linked glycoprotein-based glycophage display systems are subject to change depending on the purpose of the experiment.
• We use multiple routes to increase the titer of the glycophage during the experiment.

CD BioGlyco combines chemistry and molecular biology to provide a one-stop service for the construction of O-linked glycoproteins-based glycophage display systems. In addition, we also provide a Glycophage Display System Construction Service based on N-linked Glycoproteins. Please feel free to contact us to learn more about the process of constructing a glycophage display system based on O-linked glycoproteins. With our rich experience in glycophage display and glycosylation research, we will excel in all kinds of challenging tasks.