Shaking-Induced Aggregation and Flotation in Immunoglobulin Dispersions: Differences between Water and Water-Ethanol Mixtures

Abstract

Structural characterization by three complementary methods of laser diagnostics (dynamic light scattering, laser phase microscopy, and laser polarimetric scatterometry) has established that shaking of immunoglobulin G (IgG) dispersions in water and ethanol-water mixtures (36.7 vol %) results in two effects. First, it intensifies the aggregation of IgG macromolecules. Second, it generates bubbles with a size range that is different in each solvent. The aggregation is enhanced in ethanol-water mixtures because of IgG denaturation. IgG aggregates have a size of ∼300 nm in water and ∼900 nm in ethanol-water mixtures. The flotation of IgG is much more efficient in water. This can be explained by a better adsorption of IgG particles (molecules and aggregates) on bubbles in water as compared to ethanol-water mixtures. Bulk nanobubbles and their association with IgG aggregates were visualized by laser phase microscopy in water but were not detected in ethanol-water mixtures. Therefore, the nanobubble flotation mechanism for IgG aggregates acting in water is not feasible for ethanol-water mixtures.