title: Stability of helix-rich proteins at high concentrations.
abstract: A number of techniques, including circular dichroism, FTIR, front face fluorescence, and UV absorption spectrophotometries, dynamic light scattering, and DSC, were used to directly measure the colloidal and conformational stability of proteins in highly concentrated solutions. Using bovine serum albumin (BSA), chicken egg white lysozyme, human hemoglobin A0, and bovine fibrinogen as model proteins, the thermal transition temperatures of proteins in dilute and concentrated solutions were compared. At 10 degrees C, no significant differences in both secondary and tertiary structures were detected for proteins at different concentrations. When temperature was introduced as a variable, however, hemoglobin and fibrinogen demonstrated higher transition midpoints (T(m)s) in concentrated rather than in dilute solutions (deltaT(m) approximately 2-10 degrees C). In contrast, lysozyme and BSA in concentrated solutions exhibit a lower T(m) than in dilute solutions (deltaT(m) approximately 2-20 degrees C). From these studies, it appears that a variety of factors determine the effect of high concentrations on the colloidal and conformational stability of a particular protein. While the prediction of excluded volume theory is that high concentrations should conformationally stabilize proteins, other factors such as pH, kinetics, protein dynamics, and intermolecular charge-charge effects may affect the overall stability of proteins at high concentrations under certain conditions.
authors.affiliation: Department of Pharmaceutical Chemistry, The University of Kansas, 2030 Becker Drive, Lawrence, Kansas 66047, USA.
authors.forename: Jianxin
authors.initials: J
authors.lastname: Guo
authors.forename: Nicholas
authors.initials: N
authors.lastname: Harn
authors.forename: Aaron
authors.initials: A
authors.lastname: Robbins
authors.forename: Ron
authors.initials: R
authors.lastname: Dougherty
authors.forename: C Russell
authors.initials: CR
authors.lastname: Middaugh
mesh_terms: D000818:Animals; D002417:Cattle; D006801:Humans; D017433:Protein Structure, Secondary; D011506:Proteins / Q000737:chemistry*; D012996:Solutions / Q000737:chemistry; D013057:Spectrum Analysis; D044366:Transition Temperature; D014364:Tryptophan / Q000032:analysis
chemical_list: D011506:Proteins; D012996:Solutions; D014364:Tryptophan