title: Factors influencing antibody stability at solid-liquid interfaces in a high shear environment.
abstract: A rotating disk shear device was used to study the effect of interfacial shear on the structural integrity of human monoclonal antibodies of IgG4 isotype. Factors associated with the solution conditions (pH, ionic strength, surfactant concentration, temperature) and the interface (surface roughness) were studied for their effect on the rate of IgG4 monomer loss under high shear conditions. The structural integrity of the IgG4 was probed after exposure to interfacial shear effects by SDS-PAGE, IEF, dynamic light scattering, and peptide mapping by LC-MS. This analysis revealed that the main denaturation pathway of IgG4 exposed to these effects was the formation of large insoluble aggregates. Soluble aggregation, breakdown in primary structure, and chemical modifications were not detected. The dominant factors found to affect the rate of IgG4 monomer loss under interfacial shear conditions were found to be pH and the nanometer-scale surface roughness associated with the solid-liquid interface. Interestingly, temperature was not found to be a significant factor in the range tested (15-45 degrees C). The addition of surfactant was found to have a significant stabilizing effect at concentrations up to 0.02% (w/v). Implications of these findings for the bioprocessing of this class of therapeutic protein are briefly discussed.
authors: The Advanced Centre for Biochemical Engineering, University College London, UK.
authors: James G
authors: JG
authors: Biddlecombe
authors: Graeme
authors: G
authors: Smith
authors: Shahid
authors: S
authors: Uddin
authors: Sandrine
authors: S
authors: Mulot
authors: David
authors: D
authors: Spencer
authors: Chris
authors: C
authors: Gee
authors: Brendan C
authors: BC
authors: Fish
authors: Daniel G
authors: DG
authors: Bracewell
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000737:chemistry*; D009994:Osmolar Concentration; D010316:Particle Size; D011136:Polysorbates / Q000737:chemistry; D055550:Protein Stability; D011506:Proteins / Q000737:chemistry*; D033081:Shear Strength; D013499:Surface Properties; D013696:Temperature
chemical_list: D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D011136:Polysorbates; D011506:Proteins