CD BioGlyco is a biotechnology company that provides custom services for glycophage display-based antibody development. Our glycophage display for antibody development is a versatile, in vitro selection technology that can be utilized to discover high-affinity antibodies specific to a wide variety of glycoprotein antigens. We understand the importance of efficiently developing monoclonal antibodies (mAbs), and we are dedicated to helping you achieve that as quickly as possible via our Glycophage Display services.
Our glycophage display technology is an experimental method used in the development of antibodies, which are proteins that can recognize and bind to specific molecules. The glycophage display-based antibody development process begins with glycophage construction, followed by a panning strategy, culminating in an analysis of antibodies. This approach is highly regarded for its high throughput, selectivity, and ease of use. By harnessing the unique characteristics of the glycophage, we enhance our understanding and utilization of various biomolecules. This display system finds broad applications in drug development, antibody screening, carbohydrate-protein interaction studies, and more.
Fig.1 The process of antibody development based on glycophage display. (CD BioGlyco)
We use the M13 bacteriophage to set up glycophage display systems that enable us to bind phenotypes of interest to the glycophage surface, thereby displaying their distinct structural domains or functionalities. We leverage the periplasmic co-localization of this protein modification and phage assembly to establish a novel genetic system of glycosylation based on Escherichia coli. This allows us to generate glycophage populations that display an N-linked glycan or O-linked glycan on their surface. By locating the gene of interest on the phagemid, we could physically link the phenotypic display of glycans to most of the genes encoding components of the glycan biosynthetic pathway. We have constructed two kinks of the glycophage display system: the N-linked Glycoprotein-based Glycophage Display System and the O-linked Glycoprotein-based Glycophage Display System.
Subsequently, we employ a biopanning strategy to screen antibodies that can specifically bind to the glycophages we create. We have four types of antibody libraries to screen, including naïve library, immune library, semi-synthetic library, and synthetic library. Our extensive library collection gives us the flexibility to offer a diverse range of formats, ensuring that we can meet the specific needs of various applications.
The naïve library is created by amplifying natural sources, such as primary B-cells from non-immunized donors. This type of library can be utilized for a vast array of antigens, as one library contains a diverse collection of antibodies.
The immune library is constructed by amplifying the B-cell antibodies repertoire from immunized or previously exposed donors, allowing them to target a specific set of antigens.
The synthetic library is generated through the use of computational in silico design and gene synthesis techniques, enabling precise and controlled definitions of complementarity-determining regions (CDRs) design and composition.
The semi-synthetic library is composed of both natural sources, in which CDRs are obtained, and defined parts that are designed through in silico methods.
Fig.2 Four types of antibody libraries. (Ponsel, et al., 2011)
After biotinylating the antibodies, we proceed to mix them with glycophage-containing phage preparations. The mixture forms an immunocomplex between the antibodies and glycophages. To capture the immunocomplex, we use streptavidin-coated beads, which are specifically bound to the antibody/glycophage immunocomplex.
The biopanning process involves multiple rounds of selection, with each round enriching the population of antibodies that bind to the target glycophage. Over time, the proportion of target-specific antibodies increases with each iteration. We perform cyclic panning, which involves multiple rounds of glycophage binding to the antibody, washing, elution, and reamplifying the glycophage binders in E. coli. Through each cycle, we select specific binders that wash non-binders away. After three or four rounds, we obtain antibodies that specifically bind to the glycophages.
To validate the qualitative selection process, we utilize quantitative immunoassays such as the enzyme-linked immunosorbent assay (ELISA), immunofluorescence, homogeneous time-resolved fluorescence (HTRF), complement fixation, agglutination, and/or precipitation. These assays allow me to assess the interactions between antibodies and antigens more quantitatively. Through those comprehensive validations, we ensure the accuracy and reliability of the results.
Our services offer exceptional flexibility, allowing us to screen for antibodies that specifically bind to target glycoprotein based-glycophage from our extensive antibody library. Additionally, we also screen for glycoproteins that exhibit selective binding to target antibodies from our comprehensive glycophage library. With these capabilities, we provide a comprehensive solution for antibody development.
Technology: Phage antibody panning
Journal: PloS one
IF: 3.7
Published: 2011
Results: This article showed that by using the F1 antigen as the screening target, scFv single-chain antibody fragments that specifically bind to Yersinia pestis could be effectively screened. Eight scFvs were selected for evaluation of their binding ability to recombinant F1 antigen and F1-positive Y. pestis. Results showed that seven out of eight scFvs were able to bind to these targets. Phage-displayed scFv were found to be easier to purify, label, and more stable compared to soluble scFv. Additionally, direct fluorescent labeling of phage-displayed scFv was used for a one-step flow cytometry assay. These findings demonstrate the potential of phage-displayed protein technologies in the development of more efficient and stable immunoassays for targeted detection of Y. pestis F1 antigen.
The development of antibodies is crucial because it enables the creation of diverse diagnostic test kits based on specific interactions required. Antibody development is pivotal in drug discovery since it allows the engineering of important drug attributes such as potency, specificity, cross-reactivity, and stability. The development of antibodies also plays a crucial role in identifying novel therapeutic targets and devising effective techniques for extracting biologically active ligands. Overall, antibody development is an essential research direction in the life sciences field.
Hybridoma technology is another method that is widely used to generate monoclonal antibodies, which are highly pure, sensitive, and specific. This method involves isolating antibody-producing B lymphocytes from mice that have been immunized with a specific antigen. These B lymphocytes are then fused with immortal myeloma cell lines to create hybridoma cell lines. These hybridoma cells are cultivated in the laboratory to produce monoclonal antibodies against the target antigen. The hybridoma technology can be implemented using either an in vivo or an in vitro approach. Due to its ability to generate high-quality monoclonal antibodies, hybridoma technology is considered an excellent method among available techniques.
With our cutting-edge technology and extensive knowledge, CD BioGlyco provides specialized solutions for the discovery and engineering of antibodies that target intricate glycan antigens. Relying on the exceptional and dependable Glycan Display Platform, we are the ideal choice for conducting pharmaceutical research and developing antibodies for combating various diseases. Don't wait to get in touch! Contact us today to learn more about how we can help you.
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