Closing the Gap: Counting and Sizing of Particles Across Submicron Range by Flow Cytometry in Therapeutic Protein Products

Quantification and size distribution characterization of subvisible particles in parenteral biopharmaceutics, present as both proteinaceous and nonproteinaceous particles in the size range from 0.1 to 100 μm, are important for biopharmaceutical industry due to their potential safety and efficacy implications. Although a number of analytical techniques are available to count and size subvisible particles, characterization of particles ≤2 μm remains a significant challenge due to technical limitations of existing particle counting instruments. In this article, we demonstrate the ability of an optimized flow cytometry system to detect and quantify size distribution of subvisible particles without additional labeling that includes the critical submicron range in biopharmaceutical formulations. In addition, these qualitative and quantitative determinations are performed in a high-throughput manner using sample volumes that allow statistically significant evaluations. This approach can be used not only to ascertain the quality of therapeutic protein products but also to evaluate numerous conditions during the screening of drug candidates and their prospective formulations.