Sialylated Lactose-based Cell-free System Construction Service

Crafting Solutions, Cell-Free with Lactose Power at CD BioGlyco

The Cell-free System is a technology for protein expression in an in vitro environment, and one of the characteristics of this technology is that it does not rely on living cells. It utilizes the translation mechanism of biomolecules extracted from cells of different species, as well as the addition of an expression template plasmid to provide the reaction substrate.

CD BioGlyco has long specialized in research in the field of biology and has delved deeply into glycobiology and the construction of cell-free systems. Relying on a high level of biological expertise, CD BioGlyco provides clients with a high-quality sialylated lactose-based cell-free system construction service.

Fig.1 Sialylated lactose-based cell-free system construction services. Fig.1 A process of constructing a cell-free system based on sialylated lactose. (CD BioGlyco)

Cell-free glycoengineering

Enzymes play an important role in the construction and application of cell-free glycoengineering. We improve the ability of cell-free systems to prepare glycan molecules and glycoproteins by performing high-throughput screening of different enzymes.

For example, endoglycosynthase can disrupt specific glycosidic bonds in glycan molecules and prepare glycoproteins by transferring pre-synthesized glycans from glycosyl donors to acceptor proteins as a whole. In this process, we use glycooxazolines as glycosyl donors for glycoconjugates to facilitate transglycosylation yields.
In addition, we construct different glycoforms by different enzyme-mediated glycan remodeling reactions. In vitro, glycosylation analyses are performed to probe different enzyme substrate specificities and activities. For example, glycosyltransferases screened in strains of Escherichia coli used for in vitro glycoengineering tend to have more stringent substrate specificity and require extended glycosylated sequence subsequences.

Sialylated lactose-based cell-free system

We center on the sialylation pathway of lactose to construct cell-free systems and expand their applications.

Cell lysis: We extract cell lysates from cells expressing sialylated lactose on the cell surface.
Biomass extraction: Depending on the client's research needs, we use purified translation elements (e.g., various types of glycosyltransferases) to obtain cell-free systems.
Ingredient addition: We transfer the sialylation glycosylation mechanism of lactose to Escherichia coli strains to enable Escherichia coli to specifically transfer glycan sites to receptor proteins, thereby controlling the expression of target glycoproteins in a cell-free system.
Protein purification: Protein purification in the cell-free system is simpler and softer than in the live cell system, and we use liquid chromatography-mass spectrometry to purify the glycoproteins obtained.

Publication Data

Technology: Cell-free synthetic biology

Journal: Frontiers in Chemistry

IF: 5.5

Published: 2020

Results: In this article, the researchers used cell-free synthetic glycobiology as a highly modular framework to research and design glycan biosynthesis and biomolecule glycosylation pathways in the cell-free domain. In the process, the researchers also designed and constructed glycosyltransferase reaction systems utilizing a cell-free approach for the predictable and controllable production of glycan molecules.

Fig.2 Biological cell-free systems for the synthesis of sugar molecules. Fig.2 Cell-free biological systems for biosynthesis of glycomolecules. (Jaroentomeechai, et al., 2020)

Frequently Asked Questions

What is meant by synthetic glycobiology?
- The term "synthetic glycobiology" has been used primarily to describe the use of protein engineering and chemical methods to re-engineer glycosyltransferase assembly lines to produce specific glycan structures. This original definition narrowly refers to the use of Golgi-resident glycosyltransferases to modify protein glycosylation within and on the surface of eukaryotic cells.
What is the difference between cell-free and cell-based systems?

Cell-free system	Cell-based system
Fast and flexible, from template to purification in as little as 6h Highly controllable and adaptable No need for live cells to tolerate cell-harmful protein expression Highly efficient expression of corresponding proteins can be achieved through targeted optimization	Wide range of applications and proven technology Suitable for high-volume expression and production of corresponding proteins Prokaryotic and yeast are easy to prevent large

Advantages

Leading technology: We have long been committed to the development of biological technology, skilled in high-throughput screening, glycoengineering, the construction of cell-free systems and other advanced technologies, and constructed advanced Chassis Cell Platforms.
Rich product line: We offer a range of Product Manufacturing and services, including cell surface glycosylation assays, glycan engineering services, Chassis Cell Development, etc. These products and services cover a wide range of life science research areas and meet different research needs.
Considerate service: We customize our experimental solutions according to the needs of our clients to ensure their satisfaction.

CD BioGlyco has won the credibility of clients with leading biological technology, innovative products, and attentive service. We always adhere to the concept of professionalism first and dedicate ourselves to providing our clients with high-quality products and services. Please feel free to contact us if you have any questions, and our staff will answer you promptly.

Reference

Jaroentomeechai, T.; et al. Cell-free synthetic glycobiology: designing and engineering dlycomolecules outside of living cells. Frontiers in Chemistry. 2020, 8.

For research use only. Not intended for any clinical use.

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