brine and 2-2-4-trimethylpentane

Self-assembly of the anionic surfactant AOT with the protein alpha-lactalbumin in isooctane/brine mixtures results in phase structures whose type, size, and shape differ considerably from those formed by the surfactant alone. Small-angle X-ray scattering was used to determine the size and shape of these structures for 5.4 < pH < 11.2 and 0.25, 0.33, and 0.4 wt % NaCl. All pH values were above the reported isoelectric point for the protein. The composition of the system (except for salt) was fixed, with 2.5 wt % surfactant in equivolume mixtures of oil and water and either 0 or 0.4 wt % protein. Under these conditions, AOT in the absence of protein always formed spherical, water-in-oil (w/o) microemulsion droplets in the organic phase with no self-assembly in the aqueous phase. In the presence of alpha-lactalbumin, self-assembled structures were formed in both aqueous and organic phases, and the size and shape of these was tuned by both pH and [NaCl]. Protein-surfactant interaction was weakest at the most alkaline pH, with protein-free, spherical droplets forming in the organic phase and surfactant-decorated soluble protein clusters forming in the aqueous phase. As pH was decreased, protein increasingly partitioned to the organic phase and droplets became ellipsoidal and much larger in volume, with these effects enhanced at lower salt concentration. Aqueous structures were also strongly affected by pH, shifting from prolate protein/surfactant aggregates at alkaline pH to oil-in-water, oblate microemulsion droplets at neutral pH. At acidic pH and higher salt concentration, self-assembly shifted toward a third, anisotropic aqueous phase, which contained discoid bilayer structures. It is proposed that hydrophobic attraction causes association of the protein with the surfactant monolayer, and pH and [salt] tune the system via the protein by modifying electrostatic repulsion and monolayer curvature.

Topics: Dioctyl Sulfosuccinic Acid; Hydrogen-Ion Concentration; Lactalbumin; Models, Theoretical; Octanes; Salts; Scattering, Small Angle; Surface-Active Agents

The stability and structure of protein-containing water-in-oil (w/o) microemulsions were investigated by using the large protein immunoglobulin G (IgG, MW 155,000) in a mixture comprised of brine, sulfosuccinic acid bis [2-ethylhexyl]ester (sodium salt), and isooctane. We explored factors affecting the initial uptake of IgG into the w/o microemulsion and its subsequent release to a solid (precipitate) phase, and the kinetics of the latter process. Influences of such parameters as pH, ionic strength, and protein concentration on the solubilization and precipitation of bovine IgG in the organic phase are described. The structure and dynamics in microemulsions containing bovine IgG were probed by using dynamic light scattering, and it was found that the presence of IgG in the microemulsion induced strong attractive forces between the droplets. Based on results obtained by using these various experimental approaches, a model for protein solubilization and release is proposed. In this model, we propose the formation of clusters within which bovine IgG resides and which substantially slow the kinetics of protein release from the droplets to the precipitate phase.

Topics: Animals; Cattle; Chemical Precipitation; Diffusion; Dioctyl Sulfosuccinic Acid; Emulsions; Hydrogen-Ion Concentration; Immunoglobulin G; Isoelectric Focusing; Light; Magnetic Resonance Spectroscopy; Models, Chemical; Models, Theoretical; Molecular Weight; Octanes; Oils; Protein Binding; Salts; Scattering, Radiation; Solubility; Succinates; Time Factors; Water