Revealing the size, conformation, and shape of casein micelles and aggregates with asymmetrical flow field-flow fractionation and multiangle light scattering.

Casein (CN) micelles are naturally occurring colloidal protein aggregates present in a dispersed state in milk. In this paper we aim to obtain a detailed description of physicochemical properties of CN micelles over the entire size distribution using asymmetrical flow field-flow fractionation (AsFlFFF) connected to multiangle light scattering (MALS) and refractive index (RI) detection. Conclusions are drawn on the colloidal level regarding shape and conformation by comparison with models of colloidal particles. By using AsFlFFF-MALS-RI, it is concluded that the CN micelles are highly polydisperse with an average rms radius and hydrodynamic radius of 177 and 116 nm, respectively. The results show that the majority of CN micelles have a spherical shape, whereas a low concentration exists of larger and elongated aggregates. By comparison with models of aggregates of colloidal particles, the aggregates are shown to be anisotropic, e.g., aggregating linearly (threadlike) or in a sheet, rather than forming randomly spherical clusters. The results show that the characterization of colloidal dispersions with AsFlFFF-MALS-RI and the comparison with theoretical models are of a general character and, thus, of fundamental importance for colloidal dispersions.