Glycophag Display-based Glycoarray Service
Client Satisfaction with Glycophag Display-based Glycoarray Service Is Our Pursuit
CD BioGlyco is expanding the application of Glycophage Display by designing and packaging glycophage systems and other methods. With our very well-established Glycan Display Platform, we offer glycoarray services based on glycophages. This technology is an alternative approach to glycan arrays that effectively optimizes the production process of glycan libraries. It utilizes the biosynthetic potential of Escherichia coli to covalently link glycans to phage particles. Also, the glycan display phage obtained based on this method is easy to recover without laborious purification techniques.
Fig.1 Process of glycophag display-based glycoarray. (CD BioGlyco)
This approach generates the displayed glycan by biosynthesis in engineered E. coli cells. First, we construct engineered E. coli so that it carries a phage that encodes a phage shell protein-receptor protein fusion and a plasmid that encodes the glycan biosynthesis pathway. In the presence of glycosyltransferase from Campylobacter jejuni, the glycan is displayed on the surface of the phage particle by covalently attaching it to the fusion protein. Afterward, we verify the effectiveness of this microarray by testing the binding to the protein.
Our experimental procedure includes the following steps:
- Phagemid construction
The phage is modified and cloned to obtain phagemid particles that combine the advantages of phage and plasmid. The phages used are verified by DNA sequencing. Plasmids for glycan biosynthesis are also constructed. - Production and purification of glycophage particles
Plasmids for glycan biosynthesis and E. coli monoclones carrying phage particles are inoculated into the culture medium for cultivation. Phage particles are recovered from the culture medium and the concentration of glycophage is assayed. - Validation of glycophage
We detect the glycophages by western blot and enzyme-linked immunosorbent assay (ELISA). - Fabrication of microarrays
Phage display glycan libraries are constructed using slides as solid supports. - Microarray scanning
We are equipped with a state-of-the-art microarray scanner to scan slides and analyze experimental results.
Publication Data
Technology: Glycophage display
Journal: Biotechnology Journal
IF: 3.781
Published: 2015
Results: In this study, a new glycan array technique was constructed by combining the display of glycans on filamentous phage with the immobilization compatibility of phage particles. This technique detects the interaction between glycans and glycan-binding proteins such as lectins by modifying sugar phages with glycans of different structures. To validate this technique, this study attempted to display C. jejuni glycans from the phage surface. The results showed that these glycans were easily captured by lectins or detected by glycan-specific serum antibodies.
Fig.2 Glycan structures used in array fabrication. (Çelik, et al., 2015)
Applications
- The glycophag display-based glycoarray is used to study the molecular interactions in which glycans are involved, thus providing data to support the study of the development, growth, function, and survival of organisms.
- The glycophag display-based glycoarray is used to identify specific microbial polysaccharides that interact with human antibodies.
- The glycophag display-based glycoarray is used to determine the glycan-binding specificity of glycan-binding proteins.
- The glycophag display-based glycoarray is used to reveal key mechanistic details of glycan immunorecognition and host-pathogen interactions.
Advantages of Us
- We analyze the interactions between glycans and glycan-binding proteins using only minimal biological materials.
- We realize the full potential of the glycophag display-based glycoarray by developing reliable and reproducible chemistry and by increasing the diversity and number of glycan structures.
- Our redesigned glycophage system allows for very high phage titers.
CD BioGlyco is skilled in the chemistry associated with the synthesis, harvesting, and immobilization of glycans, which is critical to the development of glycoarray technology. We aim to expand the diversity of glycan libraries while optimizing glycan array technology to support the development of existing tools for high-throughput analysis of glycoprotein interactions. Please feel free to contact us to learn more about the experimental procedures and other information about the glycophag display-based glycoarray.
Reference
- Çelik, E.; et al. Glycoarrays with engineered phages displaying structurally diverse oligosaccharides enable high-throughput detection of glycan–protein interactions. Biotechnology Journal. 2015, 10(1): 199-209.
