Mass Detection for Peptides and Glycans in Biopharma

To learn more about how the ACQUITY QDa brings greater sensitivity, selectivity, and productivity to these routine bioipharmaceutical applications and others, please see the following presentation, or contact your local Waters representative. 

The high costs associated with meeting regulatory guidelines for biotherapeutic drug safety has renewed interest in efficient methods that reduce costs and increase productivity in the manufacturing process.

The ACQUITY QDa offers the ability to employ mass spec analysis as an orthogonal detection technique in a single workflow for peptide and glycan analyses, greatly improving monitoring capabilities in process development and production environments with minimal cost and effort.

With a plug-and-play design, and integrated compliant data acquisition and analysis through Empower 3 Software, the ACQUITY QDa is a powerful tool that can increase productivity and confidence in data analysis within the biotherapeutic production environment.

To demonstrate the utility of the ACQUITY QDa Detector for monitoring peptides, we analyzed a tryptic map of a monoclonal antibody (mAb). As shown, we analyzed this digest with eluents containing both TFA and FA and in each case we were able to detect the resulting peptides from the digest.

While there is a difference in absolute intensity, the data shows the ACQUITY QDa Detector is compatible with both TFA and FA separations. Furthermore, with additional data interrogation, we routinely demonstrate greater than 90 percent peptide map coverage, and can see multiple charge states for each peptide.

Here we show an example of released N-glycan detection, accomplished using the RapiFluor-MS labeling reagent during sample preparation, and using both a fluorescence detector and the ACQUITY QDa mass detector during analysis. Shown are the combined chromatograms of released N-glycans from three different samples chosen to reflect the wide range of glycans typically seen and monitored in biotherapeutic development and production settings. These are IgG, which includes simple bi-antennary glycan structures; RNnase B, which includes many high mannose structures; and Bovine Fetuin, which includes many larger, more complex and highly sialylated glycan structures. As you can see, both chromatograms show equivalent detection responses, and all the peaks in one are seen in the other.

Thus the ACQUITY QDa Detector provides laboratories that are monitoring glycans with increased confidence and greater productivity, as peak assignments and glycan structures can be confirmed via fluorescent response and orthogonal mass data captured within a single run.