Glycophage Display-based Epitope Mapping Service

Glycophage Display-based Epitope Mapping Service

Glycophage Display-based Epitope Mapping Service at CD BioGlyco

CD BioGlyco provides Glycophage Display-based Technology that holds significant application in the field of epitope identification and selection. Using the glycophage display method to map is an efficient approach, allowing for the rapid identification and mapping of epitopes, enabling clients to understand the underlying biology and develop targeted therapies more efficiently.

Epitope mapping is an indispensable method that allows us to delve into the intricate workings of antibodies, ensuring a comprehensive understanding of their functionality. It plays a pivotal role in advancing the fields of diagnostics and therapeutics identifying and characterizing specific antibody binding sites.

At CD BioGlyco, we use this technique for screening large libraries of glycoproteins that target a specific protein. By our method, antibodies meticulously choose glycoproteins that exhibit sufficient affinity for their combining sites. As a result, it enables the precise identification of particular epitopes. After the identification of these epitopes, they can be utilized in the development of vaccines that specifically target the particular pathogen. This targeted approach ensures that the vaccine is designed to combat the pathogen and enhance immune responses against it effectively. We also provide related services for vaccine development based on these epitopes through our Glyco™ Vaccine Development Platform.

Fig.1 General strategy from epitope selection to phage-based vaccine production. (Palma, 2023)Fig.1 General strategy from epitope selection to phage-based vaccine production. (Palma, 2023)

  • We have constructed glycophage display systems utilizing a genetically engineered bacteriophage that can present a diverse range of glycoproteins on its surface. Our system construction process involves carefully choosing a bacteriophage capable of infecting commonly used laboratory bacteria, such as Escherichia coli. M13 bacteriophage is a frequently employed choice for phage display experiments at CD BioGlyco. Our glycophage display systems come in two types, N-linked Glycoprotein and O-linked Glycoprotein. These libraries are valuable for pinpointing epitopes that recognize particular structural motifs, domains, or subdomains, as well as facilitating screening for ligands that interact with specific receptors.
  • Then we perform binding experiments with the enzyme-linked immunosorbent assay (ELISA). Glycophages are incubated with the immobilized antibodies or receptors. Antibodies or proteins of interest are exposed to a glycoprotein-based glycophage display system during incubation.
  • Through a series of washes and selection steps, we screen and retain glycophages that interact with the target antibodies or target proteins, while eliminating those that do not bind. After screening, we incubate the phages that bound to the target and determine specific epitopes on the target antibodies or target proteins.
  • To obtain epitopes with better affinity and specificity for the target antibodies, we will create a new library based on the selected glycophages and conduct another round of selection. Once potential hits are identified and optimized, we will perform comprehensive characterization. This involves assessing the binding of antibodies to the hits, evaluating their ability to induce an immune response, and determining the effectiveness and safety of the immune response. This characterization process includes biochemical or biophysical analyses, as well as in vivo studies.

Fig.2 The process of epitope mapping. (CD BioGlyco)Fig.2 The process of epitope mapping. (CD BioGlyco)

Besides, antibodies that target antigens can have multiple specificities, which means they are capable of binding to different locations on the antigens. As a result, they can have varying effects on the interaction between the antibody and antigen. We also perform a molecular dissection of the mixture of antibody specificities to research the active immunotherapy strategies, which target multiple epitopes within each antigen through complex humoral responses.

Publication Data

Technology: Screening phage display libraries, Computer modeling

Journal: Journal of Biological Chemistry

IF: 5.486

Published: 2007

Results: The study investigated the potential of phage-displayed peptide mimics of Vibrio cholerae lipopolysaccharide (LPS) as an alternative approach for the development of a cholera vaccine. Six phage clones were identified from panning with two protective monoclonal antibodies, S-20-4 and A-20-6, which target the O-antigen of V. cholerae O1 Ogawa serotype. These phage-displayed peptide mimics were able to compete with LPS for binding to S-20-4, indicating that they potentially mimic important conformational epitopes of the Ogawa antigen, rather than just being recognized functionally by S-20-4. To predict the mode of interaction between these peptide mimics and S-20-4, computer modeling was performed, and one of the highest-affinity binders, 4P-8, was selected for further study. This peptide mimic shared a similar motif with other mimics and was predicted to emulate a hairpin structure that partially mimics the interaction between Ogawa O-antigen and S-20-4, forming a unique ligand interaction with S-20-4. Furthermore, immunization of mice with 4P-8-KLH conjugate induced the production of anti-LPS antibodies. However, the immune response alone was unable to confer protection against cholera or kill V. cholerae. Nevertheless, boosting with 4P-8-KLH after LPS priming prolonged the LPS-specific IgG and IgM antibody responses and provided greater protection compared to immunization with LPS alone.

Applications

  • Glycophage display-based epitope mapping can be used in diagnostic test research for improved accuracy and efficiency, such as ELISAs or lateral flow assays.
  • Glycophage display-based epitope mapping contributes to producing valuable molecules for therapeutic research.
  • Identifying the epitope targeted by antibodies allows for the development of vaccines and therapeutics, as well as improving diagnostic effectiveness and understanding of immune responses.

Advantages

  • Our glycophage display simplifies epitope selection and screening, making it a standout technique among others.
  • Our glycophage display-based epitope mapping can be applied for vaccine development, offering the flexibility of customization by selecting and optimizing specific epitopes from pathogenic organisms.
  • Vaccines can stimulate the immune system effectively by presenting carefully chosen epitopes, surpassing the limitations of traditional methods that use whole pathogens.
  • The use of glycophage display reduces the risk of side effects and insufficient immune responses associated with using inactivated or attenuated pathogens.

CD BioGlyco is a biotechnology company, offering glycophage display-based epitope mapping service. Based on our Glycan Display Platform, we help our clients accurately identify and map specific antigenic epitopes, providing valuable tools and support for disease research, drug development, and immune vaccine design. With a combination of advanced technology and stringent quality control, our research team is committed to delivering high-quality services and customized solutions to clients. No matter if you are in academia or the pharmaceutical industry, please contact us and let's collaborate to make meaningful contributions to the advancement of biomedical research.

References

  1. Palma, M. Epitopes and mimotopes identification using phage display for vaccine development against infectious pathogens. Vaccines. 2023, 11: 1176.
  2. Dharmasena, M.N.; et al. Development of peptide mimics of a protective epitope of Vibrio cholerae Ogawa O-antigen and investigation of the structural basis of peptide mimicry. Journal of Biological Chemistry. 2007, 282(46): 33805-33816.
This service is for Research Use Only, not intended for any clinical use.

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