Adopting technologies to streamline biotherapeutics characterization

The characterization of biotherapeutics is essential to assess their safety, quality, and efficacy. New and improved methods are needed to meet the growing analytical challenges of a new wave of cancer biotherapeutics that have complex structures and impurity profiles. In this presentation, demonstrated, are two technological improvements that will enable both speed and comprehensive characterization of biologics. In the first example, a demonstration on how a time-consuming mAb digestion process (> 30 min) can be benefit from ultrafast enzymatic digestion (< 1 ms) of an intact antibody using microdroplet reactions. (Anal. Chem., 2021, 93, 3997-4005) This capability become powerful for real-time analysis of antibody subunits coupling flow-injection analysis (FIA)-MS with microdroplet reactions. Rapid generation of subunits using microdroplet digestion presents an opportunity for mobility resolved structural characterization of subunits. In addition, these subunits can be further characterized with both collision-induced dissociation (CID) and electron capture dissociation (ECD). In the second example, a workflow for profiling impurities using capillary electrophoresis (CE) techniques is demonstrated. Size-based capillary electrophoresis (i.e., cSDS, GXII) is commonly used to profile and quantify impurities. However, the identification of these impurities is often ambiguous. To overcome these challenges,high-resolution CZE-MS to improve the mapping of impurities detected by routine sized-based electrophoretic methods in a single workstream are coupled. the adoption of these techniques by the biopharma community for rapid and comprehensive analysis of biotherapeutics is envisioned.

Key learning objectives

• Significant acceleration is possible in the characterization time for proteins and antibodies.
• Accelerated microdroplet reactions for molecule characterization are possible on the Agilent mass spectrometers with the Jetstream source.
• More accurate size based analysis calibrations are possible with calibration corrections based on CE/MS determined masses.

Who should attend?

• People who need to perform or manage people who perform characterization of antibodies and protein based biotherapeutics.