title: Effect of cleaning agents and additives on Protein A ligand degradation and chromatography performance.
abstract: Protein A chromatography, employing the recombinant Protein A ligand, is widely used as a capture step for antibody and Fc-fusion proteins manufacture. Protein A ligands in these matrices are susceptible to degradation/loss when exposed to cleaning agents such as sodium hydroxide, resulting in loss of capacity on reuse. In this study, MabSelect Protein A ligand and MabSelect SuRe Protein A ligand were chosen to evaluate the impact of alkaline cleaning solutions on the ligands and the packed columns. The Protein A ligands alone and the Protein A columns were incubated or cycled in different concentrations of sodium hydroxide solutions with and without additives, respectively. Ligand integrity (degradation) and ligand function (binding affinity) were studied using SDS-PAGE and customized Biacore technology, surface plasma resonance (SPR) and were successfully correlated with column performance measurement in terms of static binding capacity (SBC), dynamic binding capacity (DBC) and recovery as a function of exposure to cleaning agents with and without additives. The findings and the methodology presented in this study are not only able to determine appropriate cleaning conditions for Protein A chromatography, but also provided tools to enable systematic and rapid study of the cleaning solutions and conditions.
authors: Manufacturing Sciences, AbbVie Bioresearch Center, Worcester, MA 01605, USA. Electronic address: lihua.yang@abbvie.com.
authors: Lihua
authors: L
authors: Yang
authors: Manufacturing Sciences, AbbVie Bioresearch Center, Worcester, MA 01605, USA. Electronic address: jason.harding@abbvie.com.
authors: Jason D
authors: JD
authors: Harding
authors: Global Biologics, AbbVie Bioresearch Center, Worcester, MA 01605, USA. Electronic address: ivanov.alexander@abbvie.com.
authors: Alexander V
authors: AV
authors: Ivanov
authors: Manufacturing Sciences, AbbVie Bioresearch Center, Worcester, MA 01605, USA. Electronic address: natraj.ram@abbvie.com.
authors: Natarajan
authors: N
authors: Ramasubramanyan
authors: Manufacturing Sciences, AbbVie Bioresearch Center, Worcester, MA 01605, USA. Electronic address: diane.dong@abbvie.com.
authors: Diane D
authors: DD
authors: Dong
mesh_terms: D000906:Antibodies / Q000378:metabolism; D002846:Chromatography, Affinity / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D008024:Ligands; D011994:Recombinant Proteins / Q000378:metabolism; D012972:Sodium Hydroxide / Q000737:chemistry; D013205:Staphylococcal Protein A / Q000378:metabolism
chemical_list: D000906:Antibodies; D008024:Ligands; D011994:Recombinant Proteins; D013205:Staphylococcal Protein A; D012972:Sodium Hydroxide
keywords: Binding capacity; Cleaning solution; Monoclonal antibody; Protein A chromatography; Protein A ligand; Surface plasma resonance