The Effective Production of Mono- or Di- Fucosylated Lactose Based on Cell-free System Construction at CD BioGlyco

At CD BioGlyco, our service encompasses a comprehensive approach to the synthesis of mono- or di-fucosylated lactose structures within a Cell-free Environment. We leverage advanced techniques to select and incorporate appropriate glycosyltransferases (GTs) into the construction process. By screening α1,3 and α1,2 GTs from H. pylori and designing pathways with the suitable GTs, we ensure the precise assembly of glycosylation pathways tailored to our clients' needs.

Our service involves the synthesis of enzymes through separate cell-free protein synthesis (CFPS) reactions, followed by the strategic combination with suitable sugar donors to initiate in vitro glycosylation (IVG) reactions. Through meticulous optimization of reaction conditions, we strive to enhance the efficiency and yield of mono- or di-fucosylated lactose synthesis. Based on our cell-free system, we provide a powerful Cell-based Production Chassis Library, and clients also can choose the interested chassis cell and work together to Develop the Custom Chassis.

Fig.1 The process of Mono- or Di- fucosylated lactose-based cell-free system construction. (CD BioGlyco)

• Select the GTs involved in the fucosylation of lactose. In this case, it needs to select α1,3 and α1,2 fucosyltransferases.
• Design the biosynthetic pathway by combining the selected GTs with the necessary enzymes for lactose synthesis. In this case, it needs enzymes such as ApNGT (N-glycosyltransferase) and NmLgtB (lactose synthase).
• Express and optimize the soluble expression of each GT using CFPS, which involves expressing the GT genes in a CFPS system and optimizing the conditions for high yield and solubility.
• Synthesize the enzymes and GTs for glycan extension in separate CFPS reactions, which involves expressing the enzymes and GTs required for lactose synthesis separately in CFPS reactions.
• Mix the CFPS products containing the enzymes and GTs with appropriate sugar donors to form IVG reactions, which involves combining the CFPS products with the necessary sugar donors for fucosylation.
• Purify the IVG products and analyze them using mass spectrometry (MS) to confirm the presence of fucosylated lactose.

Applications of Mono- or Di- Fucosylated Lactose-based Cell-free System

• The mono- or di-fucosylated lactose-based cell-free system can be employed to produce glycoproteins with tailored glycan structures that mimic natural antigens, facilitating the development of novel immunotherapies and vaccines. Clients also can find our pre-produced Glycan Products on our website.
• The synthesized mono- or di-fucosylated lactose structures serve as valuable standards for glycan profiling and analysis. Researchers can use these structures as reference materials for characterizing glycan composition, abundance, and modifications in biological samples, aiding in the study of various diseases and biological processes.
• The mono- or di-fucosylated lactose-based cell-free system offers a streamlined approach for the production of recombinant glycoproteins with defined glycan structures.

Advantages

• At CD BioGlyco, we uphold stringent quality control measures at every stage of the cell-free system construction process.
• At CD BioGlyco, our services encompass the entire spectrum of mono- or di-fucosylated lactose-based cell-free system construction, from enzyme selection and optimization to reaction design and analysis.
• At CD BioGlyco, our team is dedicated to staying abreast of the latest advancements in glycosylation pathways, ensuring that clients benefit from cutting-edge solutions.

Similar Services at CD BioGlyco

CD BioGlyco combines expertise, innovation, and commitment to excellence to deliver unparalleled advantages in mono- or di-fucosylated lactose-based cell-free system construction. With CD BioGlyco as your partner, you can navigate the complexities of glycoscience research with confidence and success. Please feel free to contact us to learn more about our services.

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