How does the automated oligosaccharide synthesis system improve upon traditional chemical methods?

The automated system utilizes enzymatic glycosylation, offering high regio- and stereo-specificity under mild reaction conditions, eliminating the need for extensive protection and deprotection steps required in chemical glycosylation. A temperature-dependent polymer, specifically poly(N-isopropylacrylamide) (PNIPAM)is used, that acts as a soluble support for the oligosaccharide during synthesis. The entire process is controlled by a CEM Liberty Blue peptide synthesizer, which automates the addition of reagents, temperature control, and washing steps.

What role does the cleavable linker play in the automated oligosaccharide synthesis, and what type is used?

The automated system uses a cleavable linker, specifically a thioether linker, to connect the oligosaccharide primer to the PNIPAM polymer. This linker is stable during enzymatic glycosylation but can be cleaved under mild oxidation with hydrogen peroxide to release the free oligosaccharide, avoiding harsh conditions that could damage the sensitive glycan molecules.

What challenges were encountered when trying to apply Merrifield's concept to enzymatic glycosylation within this automated system?

The system initially attempted to use Merrifield's concept for enzymatic glycosylation. However, enzymatic synthesis using a solid-phase method faced challenges due to enzyme incompatibility with the solid resin. This incompatibility hindered the desired results, leading to the adoption of a soluble support strategy using PNIPAM.

What are some potential future improvements planned for the automated glycan synthesis system?

Future improvements to the automated system include expanding storage capacity for reagents and enzymes, integrating real-time monitoring capabilities to track reaction progress, and creating a comprehensive enzyme kinetics database to optimize reaction conditions. These enhancements promise even greater efficiency and versatility in automated glycan synthesis.

Beyond accelerating research, what are the broader implications of automated oligosaccharide synthesis for medicine and healthcare?

Oligosaccharides play key roles in various biological and medical applications, including diagnostics and vaccine development. The ability to synthesize them efficiently and in large quantities accelerates research in these areas, potentially leading to faster drug discovery, improved diagnostics, and advancements in personalized medicine, ultimately improving patient outcomes and healthcare strategies.

Automated machine synthesizing sugar molecules, representing the intersection of technology and biology in glycan synthesis.

Automated Chemistry: How Machines are Revolutionizing Oligosaccharide Synthesis

Avery Sinclair in Tech & Innovation September 2025 • 4 min read.

"A new machine-driven enzymatic system promises to accelerate research and development in glycobiology and medicine."

Oligosaccharides, vital components in biology, play key roles in many biological and medical applications, from diagnostics to vaccine development. Obtaining sufficient quantities of pure oligosaccharides for research has been a major bottleneck. Traditional methods are time-consuming and labor-intensive, which underscores the need for automation. In a breakthrough study, researchers introduce a fully automated system for synthesizing oligosaccharides using enzymatic glycosylation.

Inspired by advancements in automated oligonucleotide and oligopeptide synthesis, this new system leverages a commercially available peptide synthesizer. Unlike chemical glycosylation, which requires extensive protection and deprotection steps, enzymatic glycosylation offers high regio- and stereo-specificity under mild reaction conditions. This automated approach represents a significant leap forward, potentially democratizing access to complex glycans for researchers worldwide.

The innovative system combines enzymatic reactions, a temperature-dependent polymer, and a peptide synthesizer to streamline the synthesis process. This method simplifies the creation of oligosaccharides and makes it more accessible to researchers without specialized expertise.

Unlocking the Power of Automation: Key Innovations in Oligosaccharide Synthesis

Automated machine synthesizing sugar molecules, representing the intersection of technology and biology in glycan synthesis.

The automated system is based on a thermosensitive polymer, poly(N-isopropylacrylamide) (PNIPAM), which acts as a soluble support for the oligosaccharide during synthesis. At temperatures below the lower critical solution temperature (LCST), PNIPAM is soluble in water, allowing enzymes to access the growing glycan chain. When the temperature increases above the LCST, PNIPAM becomes insoluble, precipitating out of the solution along with the attached oligosaccharide. This phase transition allows for easy separation of the product from the reaction mixture through simple filtration.

A cleavable linker connects the oligosaccharide primer to the PNIPAM polymer. This linker is designed to be stable during the enzymatic glycosylation steps but can be cleaved under mild conditions to release the free oligosaccharide. The researchers used a thioether linker, which can be cleaved by oxidation with hydrogen peroxide, avoiding harsh acidic or basic conditions that could damage the sensitive glycan molecules.

Reaction and Separation: The automated process hinges on effective reaction and separation steps for sugar elongation.
Merrifield's Concept: An initial attempt was made to use Merrifield's concept for enzymatic glycosylation; however, enzymatic synthesis using a solid-phase method didn't yield the desired results.
Enzyme Incompatibility: The incompatibility between enzymes and solid resin was a significant challenge.

The entire process is controlled by a CEM Liberty Blue peptide synthesizer, which automates the addition of reagents, temperature control, and washing steps. The synthesizer is equipped with a reaction vessel connected to a filter and a temperature-control system, all managed by a computer. Enzymes, nucleotide sugars, and reaction buffers are stored in tubes and automatically injected into the reaction vessel.

The Future of Glycan Synthesis: Accessible and Automated

This automated system offers a powerful tool for glycan research, making the synthesis of complex oligosaccharides more accessible to a broader range of scientists. By reducing the need for specialized expertise and minimizing labor-intensive steps, this technology has the potential to accelerate drug discovery, vaccine development, and our understanding of the biological roles of glycans. Future improvements, such as expanding storage capacity, integrating real-time monitoring, and creating a comprehensive enzyme kinetics database, promise even greater efficiency and versatility in automated glycan synthesis.