Constructing Innovative Cell-Free Systems with Sialylated LacNAc-Based Technology at CD BioGlyco

Cell-free Systems generally refer to the use of cell lysates, amino acids, nucleic acids, and cofactors to produce biomolecules or chemicals, in the absence of intact cells. In addition, cell-free systems are capable of designing and synthesizing glycosylation in the absence of cells using synthetic biology methods such as metabolic engineering, protein engineering, and others.

With state-of-the-art synthetic biology technology, CD BioGlyco has been working in the field of cell-free systems for several years and provides a systematic and efficient sialylated LacNAc-based cell-free system construction service to clients.

Construction of LacNAc terminated by sialic acid

We use sialyltransferase to install Sia residues into LacNAc. In this process, we select a variety of bacteria to screen for suitable sialyltransferases, such as Pasteurella multocida, Vibrio vulnificus, Phosphorobacterium, Campylobacter jejuni, and Escherichia coli.

Construction of cell-free systems

Cell-free systems accomplish transcription, translation, folding, post-modification, and energy metabolism processes in vitro by utilizing extracts from microbial, animal, and plant cells and supplementing them with substrates and energy, using exogenous mRNA or DNA as templates. At CD BioGlyco, we construct many types of cell-free systems based on sialylated LacNAc, such as:

• Extract-based system: We offer a wide variety of sialylated LacNAc sources such as Escherichia coli, yeast cells, animal cells, and more.

• Purified systems

In constructing cell-free systems, we construct cell-free systems with better performance by stimulating central metabolism. And, we utilize hydrocarbon polymers as energy substrates to reduce energy costs. Meanwhile, we use plasmids or linear polymerase chain reaction products as gene expression templates to obtain a cell-free system with good stability.

Cell-free synthetic glycosylation system

We introduce different glycosylation mechanisms into bacterial lysates using a bacterial cell-free glycosylation system. We install sialylated LacNAc on glycoproteins by overexpressing the encoded plasmid in a Bacterial Chassis strain.
We also take advantage of the flexibility and high throughput of cell-free systems to better design and optimize the sialylation pathway of LacNAc.

For example, we add protein-lipid chassis extracts to express sialyltransferase homologs in a cell-free system. Next, LacNAc is constructed stepwise from the glycan donor by cell-free system-generated enzymes. After that, the obtained LacNAc can be further modified by refined glycosyltransferases and trimmed with glycosidases (there are two major classes of glycosidases, exoglycosidases that remove glycans from the glycan termini and endoglycosidases that hydrolyze glycosidic bonds within the glycan chains).

Frequently Asked Question

• What are the uses of cell-free systems?
  • Synthesizing pharmaceutical proteins: The cell-free prokaryotic system Escherichia coli system provides a rapidly designed synthetic platform for the rapid production of vaccines and has also become an emerging system for the synthesis of antibodies.
  • Synthesis of post-modified drugs: Cell-free systems can bind non-natural amino acid sites specifically to proteins, enabling many kinds of post-modification.
  • Portable assays: Cell-free powder systems are made more stable and high-density for storage and transportation by direct extraction of lyophilized and powdered energy systems.

Advantages

• We have world-leading synthetic biology and high-throughput technologies to provide custom cell-free system construction, Chassis Cell Platform construction, and Chassis Cell Development services.
• We have an experienced team of scientists and a specialized technology platform to efficiently accomplish the Production of many types of glycosides.
• We use a combination of biological, chemical, and pharmaceutical techniques to ensure that each program is completed efficiently.

CD BioGlyco is dedicated to research in the field of biology, providing comprehensive customized cell-free system construction and application services. At the same time, we utilize a variety of expression analysis techniques to help our clients complete their research. Please feel free to contact us with your project needs or any questions you may have.

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