CD BioGlyco has established high-quality Platforms for glycobiology research. We provide clients with professional and efficient glycoengineering services, such as glyco-engineered systems for glycoprotein expression, protein glycoengineering, and antibody glycoengineering.

Overview of Glycoengineering

Almost all therapeutic proteins are glycosylated. The clinical efficacy and safety of therapeutic glycoproteins are influenced by the distribution, structure, and composition of their glycosyl groups. Glycoengineering is a method of improving protein properties by changing glycosylation. Specifically, glycoengineering is performed by changing the number and location of glycosylation sites or by changing the glycan structure of individual glycosylation sites. Glycoengineering is divided into two categories according to the produced way of glycoproteins: cell-based glycoengineering and chemical-based glycoengineering. Here, we will describe these two classes of glycoengineering in detail.

• Cell-based glycoengineering

Mammalian cells, especially Chinese hamster ovary (CHO) cells, have been widely used for the production of therapeutic glycoproteins. One glycoengineering strategy is to change the structure of glycans on proteins in CHO cells by knocking out genes. For example, Knockout of α-1,6-fucosyltransferase (FUT8) in CHO cells to produce therapeutic immunoglobulin (IgG) antibodies with enhanced antibody-dependent cell-mediated cytotoxicity (ADCC). There is also a glycoengineering strategy to improve the efficacy of therapeutic glycoproteins by changing the number of glycosylation sites. For example, the introduction of a new N-glycosylation site into human erythropoietin (hEPO) increases its clinical efficacy. In addition, many other cell types have been reported for protein glycoengineering, such as plant cells, insect cells, yeast cells, and bacterial cells.

Fig.1 Cell-based glycoengineering. (Jaroentomeechai, et al., 2020)

• Chemical-based glycoengineering

Chemical-based glycoengineering strategies are mainly catalyzed by glycosidase or glycosyltransferase to generate glycosidic bonds or cleavage of glycosidic bonds. For example, the efficacy of IgG is enhanced by changing the glycosidic linkage between the terminal sialic acid residue and the penultimate galactose residue. This strategy readily produces homogeneous proteins.

Fig.2 Endoglycosynthase (ENGases)-mediated preparation of homogeneous N-glycoproteins. (Jaroentomeechai, et al., 2020)

Key Technologies

CD BioGlyco leverages state-of-the-art technologies to manipulate and control glycosylation with precision and reproducibility. Our core technologies include sophisticated molecular biology techniques for gene editing and pathway engineering, as well as advanced enzymatic and chemical synthesis methods. We utilize specialized expression systems, including glyco-engineered mammalian and microbial cells, and even plant-based systems, to precisely control the glycan structures attached to target proteins.

Advanced Glycoengineering Services: Optimizing Glycoconjugate Functionality

CD BioGlyco offers a comprehensive range of glycoengineering services designed to meet diverse research and development needs. Our services span from modifying the cellular machinery to produce desired glycans to enzymatic manipulation of glycans on purified proteins, providing flexible and powerful tools for optimizing your biomolecules. Our glycoengineering services follow a systematic and client-centric workflow to ensure project success. Our services include:

To achieve specific glycan structures on recombinant proteins, we develop and utilize specialized expression systems with modified glycosylation pathways. This approach provides a high degree of control over the final glycan profile, ensuring homogeneity and improved functionality for your target protein.

• Protein Glycoengineering

We employ a panel of glycosyltransferases and glycosidases to precisely trim, remove, or add specific sugar residues to achieve a desired glycan profile. This approach is highly effective for post-translational modification, allowing for the fine-tuning of protein properties, such as enhanced stability, controlled immune response, or improved binding affinity.

• Antibody Glycoengineering

Glycosylation of antibodies, particularly on the Fc region, is a critical determinant of their effector functions, such as antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC). Our antibody glycoengineering service uses enzymatic methods to modify these glycans, enabling us to enhance or reduce specific immune functions. This is a vital tool for optimizing therapeutic antibodies and developing next-generation biologics with tailored efficacy.

Cell line glycoengineering involves the stable genetic modification of a host cell to produce glycoproteins with a desired, uniform glycan profile. Unlike transient methods, this approach permanently alters the cell's glycosylation machinery, ensuring consistent and reproducible protein production with a specific glycoform. We use advanced gene-editing tools to precisely knock out or introduce key glycosyltransferase genes. This allows for the creation of a "designer" cell line that eliminates unwanted glycans or synthesizes novel ones, providing a robust platform for biopharmaceutical manufacturing.

Sialic acid is a critical terminal glycan that significantly impacts a glycoprotein's half-life and biological activity. Our specialized services enable precise modification of sialic acid residues, using various precursors and enzymatic approaches to optimize this key glycan for improved function.

This service provides a powerful method to dynamically and specifically alter the glycan profile on the surface of living cells. We utilize various strategies, including metabolic glycoengineering, where cells are fed with engineered sugar precursors that are then incorporated into their glycans. This technique allows for the site-specific introduction of "tag-able" functional groups, which can be used for cell labeling with fluorescent probes, drug delivery systems, or for engineering cells with enhanced targeting capabilities. Our team also employs enzymatic and chemoenzymatic methods to precisely add or remove specific glycan structures from the cell surface, providing a highly controlled approach for functional modulation.

• Cell Surface N-glycan Glycoengineering

This service targets the asparagine-linked glycans (N-glycans) present on cell surface proteins. By manipulating the cellular machinery or using metabolic engineering with sugar analogs, we modify the N-glycan structures to study their roles in protein folding, quality control, and cell adhesion. This is particularly important for research into cancer and viral infections, where N-glycan profiles are often altered, and for the development of cell-based therapies with improved targeting or reduced immunogenicity.

• Cell Surface O-glycan Glycoengineering

O-glycans are attached to serine or threonine residues of cell surface proteins and are crucial for cellular signaling, differentiation, and motility. Our O-glycan glycoengineering service allows for the controlled alteration of these structures. This enables researchers to investigate the impact of O-glycans on key biological processes and to create cells with modified adhesive properties, which is valuable in tissue engineering and regenerative medicine.

• Cell Surface GPI Anchor Glycoengineering

GPI anchors are complex glycans that tether a variety of proteins to the outer leaflet of the cell membrane. Our GPI anchor glycoengineering service offers methods to modify these anchors, allowing for the precise attachment or release of specific proteins from the cell surface. This is a powerful tool for studying protein function, engineering cell-based drug delivery systems.

• Cell Surface GAG Glycoengineering

GAGs are long, unbranched polysaccharides that form a major component of the extracellular matrix and are vital for cell communication and tissue structure. This service focuses on modifying the GAG layer on the cell surface using metabolic or enzymatic approaches. By altering the composition and charge of GAGs, we modulate cellular interactions, cell signaling, and the overall microenvironment, which is highly relevant for research in tissue repair and disease progression.

• Cell Surface Glycolipid Glycoengineering

Glycolipids are essential components of the cell membrane, with their glycan head groups serving as recognition sites. Our glycolipid glycoengineering service enables the modification of these glycans, providing a means to study their roles in cell-cell recognition, signal transduction, and disease processes.

• Cell Surface Glycoprotein Glycoengineering Service

This comprehensive service offers a holistic approach to modifying the entire population of glycoproteins on the cell surface. By combining various techniques, including metabolic engineering and chemoenzymatic modification, we achieve a broad alteration of the cell's glycome. This is especially useful for creating new cellular products for therapeutic applications, such as engineered CAR-T cells with enhanced persistence, or for developing universal donor cells by masking immunogenic surface markers.

• Cell Surface Blood Group Antigens Glycoengineering

Blood group antigens are specific glycans found on the surface of red blood cells and other cell types. Our glycoengineering service is used to alter or remove these antigens, offering new possibilities for creating "universal" cells that are safely used in transfusion medicine and transplantation.

• Cell Surface Sialic Acid Glycoengineering

Sialic acids are critical terminal sugar residues that cap many cell surface glycans and are central to modulating cell recognition, immune responses, and the circulatory half-life of cells. Our specialized service provides precise control over the level and type of sialic acid expression. By adding or removing sialic acid residues, we significantly alter a cell's fate in the body, which is a key strategy for developing stealth cells that evade immune detection or for creating more effective cellular therapies.

Workflow

Publication Data

Applications

• Our solutions are particularly valuable in the biopharmaceutical industry for developing novel vaccines, monoclonal antibodies, and therapeutic enzymes with enhanced efficacy and safety.
• In immunology and oncology, glycoengineering is used to modulate immune responses by altering glycans on the surface of cells, enabling the design of next-generation cellular therapies like CAR-T cells with improved persistence or stealth cells that evade immune detection.
• Our services can be used to develop more specific and sensitive assays for the detection of diseases by engineering probes that recognize unique glycan biomarkers.
• Glycoengineering also plays a critical role in material science, where it is applied to design biomaterials with enhanced biocompatibility and controlled cellular interactions, such as those used in tissue engineering and implantable devices.

Advantages

• We offer unmatched precision in glycan modification, leading to therapeutic candidates with improved efficacy, reduced immunogenicity, and extended half-life.
• Our robust processes ensure exceptional batch-to-batch consistency and high yields, which are crucial for both research and commercial applications.
• Our custom-tailored approach, coupled with our state-of-the-art analytical capabilities, provides clients with a seamless and reliable path to optimized glycoconjugates.

Frequently Asked Questions

At CD BioGlyco, we are committed to being your partner in glycobiology. Our comprehensive glycoengineering services are designed to advance your research and development, providing you with high-quality, customized solutions. Our team of experts is ready to assist you in designing and executing the perfect glycoengineering strategy to achieve your project goals. Please feel free to contact us for more information and to discuss your project.

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