During the manufacturing of a biotherapeutic, a processor must track many features, including glycans. These molecules impact the shape and function of proteins, including their binding characteristics, stability, and solubility.
“Glycosylated proteins, also known as glycoproteins, are involved in a variety of important biological processes,” says Nigel Skinner, head of marketing at the Andrew Alliance, a Waters company. “Many recombinant biopharmaceuticals are glycoproteins, and glycosylation is routinely monitored for the development of therapeutically safe and effective biologics.”
During bioprocessing, “the host cell’s biosynthesis of N-glycans is routinely monitored, as their composition and structure are crucial for the biological activity of the drug,” explains Skinner. That’s crucial, because even “slight changes in manufacturing conditions can alter the glycosylation patterns of recombinant proteins and consequently the biological activity, safety, stability, efficacy, and immunogenicity of the end drug product,” he says.
Given the fundamental nature of those impacts of glycans, tracking them is crucial in bioprocessing, but it’s not easy. “During the process development and manufacturing of biopharmaceutical products, researchers may require screening many samples to identify those likely to have a desired, precise glycan profile,” Skinner continues. “Consequently, glycan sample preparation can be time-consuming and monotonous.”
The glycans also resist some forms of analysis, including mass spectrometry (MS). For example, “Glycans are not optically active in their native state, and also difficult to ionize in MS analysis, therefore sample labeling is required to increase fluorescence and MS response,” notes Steven Calciano, senior product manager at Waters.
Also, bioprocessors often prepare the samples manually, but it must be reproducible. According to Skinner, “Robust and reproducible methods for glycan preparation and analysis are a prerequisite for highly regulated glycoprotein-based biotherapeutics to ensure accuracy and consistency.”
He also describes a way to solve that challenge: “The end-to-end RapiFluor-MS N-glycan sample preparation using the Andrew+ pipetting robot provides scientists with the required components to successfully perform fast and efficient N-glycan release, labeling, and clean-up while reducing manipulation time for multiple samples with the use of Andrew Alliance electronic 8-channel pipettes,” he says.
It’s also possible to add more automation to glycan analysis. “The GlycoWorks Andrew+ workstation enables accurate and precise liquid handling throughout the experiment,” Skinner explains. “All protocol steps are carried out on OneLab, which records the actual execution of the protocol, facilitating the identification of potential errors.”
These steps play crucial roles in ensuring effective biopharmaceuticals. Improving the process for manufacturing treatments depends on more control during bioprocessing.