title: Linking the solution viscosity of an IgG2 monoclonal antibody to its structure as a function of pH and temperature.
abstract: Although the viscosity of concentrated antibody solutions has been the focus of many recent studies, less attention has been concentrated on how changes in protein structure impact viscosity. This study examines viscosity profiles of an immunoglobulin G (IgG) 2 monoclonal antibody at 150 mg/mL as a function of temperature and pH. Although the structure of the antibody at pH 4.0-7.0 was comparable at lower temperatures as measured by second derivative UV absorbance and Fourier transform infrared spectroscopy, differences in 8-anilino-1-naphthalene sulfonate (ANS) fluorescence intensity indicated small structural alterations as a function of pH. Below the structural transition onset temperature, the viscosity profiles were pH dependent and linearly correlated with fluorescence intensity, and followed semilogarithmic behavior as a function of temperature. The transitions of the viscosity profiles correlated well with the major structure transitions at a protein concentration of 150 mg/mL. The viscosity correlated particularly well with ANS fluorescence intensity at 0.2 mg/mL below and above the structural transition temperatures. These results suggest: (1) ANS can be an important measure of the overall structure and (2) hydrophobic interactions and charge-charge interactions are the two major physical factors that contribute collectively to the high viscosity of concentrated IgG solutions.
authors.affiliation: Macromolecule and Vaccine Stabilization Center, Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, Kansas, 66047.
authors.forename: Weiqiang
authors.initials: W
authors.lastname: Cheng
authors.forename: Sangeeta B
authors.initials: SB
authors.lastname: Joshi
authors.forename: Nishant Kumar
authors.initials: NK
authors.lastname: Jain
authors.forename: Feng
authors.initials: F
authors.lastname: He
authors.forename: Bruce A
authors.initials: BA
authors.lastname: Kerwin
authors.forename: David B
authors.initials: DB
authors.lastname: Volkin
authors.forename: C Russell
authors.initials: CR
authors.lastname: Middaugh
mesh_terms: D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000737:chemistry*; D011487:Protein Conformation; D055550:Protein Stability; D013050:Spectrometry, Fluorescence; D017550:Spectroscopy, Fourier Transform Infrared; D013696:Temperature; D014783:Viscosity
chemical_list: D007074:Immunoglobulin G
keywords: FT-IR; Fluorescence spectroscopy; high concentration; monoclonal antibody; partial specific volume; absorption spectroscopy; protein; structure; viscosity; zeta potential