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We demonstrate the use of holographic video microscopy to detect individualsubvisible particles dispersed in biopharmaceutical formulations and todifferentiate them based on material characteristics measured from theirholograms. The result of holographic analysis is a precise and accuratemeasurement of the concentrations and size distributions of multiple classes ofsubvisible contaminants dispersed in the same product simultaneously. Wedemonstrate this analytical technique through measurements on model systemsconsisting of human IgG aggregates in the presence of common contaminants suchas silicone oil emulsion droplets and fatty acids. Holographic video microscopyalso clearly identifies metal particles and air bubbles. Being able todifferentiate and characterize the individual components of such heterogeneousdispersions provides a basis for tracking other factors that influence thestability of protein formulations including handling and degradation ofsurfactant and other excipients.

Quantitative differentiation of protein aggregates from other subvisible particles in viscous mixtures through holographic characterization