German researchers have developed a technique for glycan analysis using cheap, widely available chemicals. The team, from HTW University of Applied Sciences in Berlin, was interested in reducing the cost of analyzing glycosylation—a key measure of product quality for some advanced therapies.
“One of the problems prohibiting the collection of large amounts of glycosylation data is the prohibitively expensive cost of processing glycan samples,” explains Henning von Horsten, PhD, a professor of industrial biomanufacturing at HTW-Berlin.
Processing the samples currently requires enzymatic digestion, mass spectrometry, and other tedious labor-intensive processes, he adds.
Faster and less expensive
The scientists hope their technique will help investigators analyze the composition of protein-linked glycans faster and at a reduced cost-per-measure point. They believe the latter could enable more frequent in-process controls of glycan composition during production of the next generation of advanced therapies.
von Horsten had been inspired by previous publications that reported on releasing glycans using household bleach, and wondered if the underlying mechanism of glycan release could be extended to other cheap and simple chemicals. He and his team discovered they could free up N-glycans using Oxone®, a common non-chlorine oxidizing agent, and bromide, two chemicals that are commercially sold in the solid state.
von Horsten points out that solid chemicals benefit from an extended shelf-life compared to dissolved reagents, can be shipped in their concentrated form without excess weight from solvents, and enable fine-tuned dosing.
The team is currently adapting their chemical glycan release method to rapidly free up O-glycans, as well. Unlike N-glycans, which he says are easily liberated from glycoproteins by chemical release methods, O-glycan release is more difficult and challenging.
Currently, hydrazine, a highly toxic but common chemical used as a component in rocket fuel is used for O-glycan release, he explains.
“At the moment, O-glycans are a bit of an underrepresented area because no one wants to play around with hydrazine on a regular basis,” he says, noting that they hope to replace this reagent and enable O-glycan release with the help of less harmful Oxone and bromide reagents.
“The next thing to work on is doing this with automation, using a liquid handler,” he says, adding that this would aid high-throughput analysis of glycans and generate the data volumes needed to derive new information via artificial intelligence.
The team has published some of its work in Frontiers in Molecular Biosciences.