title: The limitations of an exclusively colloidal view of protein solution hydrodynamics and rheology.
abstract: Proteins are complex macromolecules with dynamic conformations. They are charged like colloids, but unlike colloids, charge is heterogeneously distributed on their surfaces. Here we overturn entrenched doctrine that uncritically treats bovine serum albumin (BSA) as a colloidal hard sphere by elucidating the complex pH and surface hydration-dependence of solution viscosity. We measure the infinite shear viscosity of buffered BSA solutions in a parameter space chosen to tune competing long-range repulsions and short-range attractions (2 mg/mL ≤ [BSA] ≤ 500 mg/mL and 3.0 ≤ pH ≤ 7.4). We account for surface hydration through partial specific volume to define volume fraction and determine that the pH-dependent BSA intrinsic viscosity never equals the classical hard sphere result (2.5). We attempt to fit our data to the colloidal rheology models of Russel, Saville, and Schowalter (RSS) and Krieger-Dougherty (KD), which are each routinely and successfully applied to uniformly charged suspensions and to hard-sphere suspensions, respectively. We discover that the RSS model accurately describes our data at pH 3.0, 4.0, and 5.0, but fails at pH 6.0 and 7.4, due to steeply rising solution viscosity at high concentration. When we implement the KD model with the maximum packing volume fraction as the sole floating parameter while holding the intrinsic viscosity constant, we conclude that the model only succeeds at pH 6.0 and 7.4. These findings lead us to define a minimal framework for models of crowded protein solution viscosity wherein critical protein-specific attributes (namely, conformation, surface hydration, and surface charge distribution) are addressed.
authors.affiliation: Formulation Sciences Department, MedImmune, Gaithersburg, Maryland.
authors.forename: Prasad S
authors.initials: PS
authors.lastname: Sarangapani
authors.forename: Steven D
authors.initials: SD
authors.lastname: Hudson
authors.forename: Kalman B
authors.initials: KB
authors.lastname: Migler
authors.forename: Jai A
authors.initials: JA
authors.lastname: Pathak
mesh_terms: D000818:Animals; D002417:Cattle; D003102:Colloids; D004058:Diffusion; D057446:Hydrodynamics; D006863:Hydrogen-Ion Concentration; D008958:Models, Molecular; D011487:Protein Conformation; D012212:Rheology; D012710:Serum Albumin, Bovine / Q000737:chemistry* / Q000378:metabolism; D013499:Surface Properties; D014783:Viscosity; D014867:Water / Q000737:chemistry
chemical_list: D003102:Colloids; D014867:Water; D012710:Serum Albumin, Bovine