Comparison of standard and new generation hydrophobic interaction chromatography resins in the monoclonal antibody purification process.
Recent efforts to improve hydrophobic interaction chromatography (HIC) for use in monoclonal antibody (mAb) purification have focused on two approaches: optimization of resin pore size to facilitate mAb mass transport, and use of novel hydrophobic charge induction (HCIC) mixed mode ligands that allow capture of mAbs under low salt conditions. We evaluated standard HIC and new generation HIC and HIC-related chromatography resins for mAb purification process efficiency and product quality both as isolated chromatography steps and in purification process trains. We find that HIC resins with optimized pore size have significantly improved binding capacity which can increase HIC purification unit operation efficiency. The HCIC Mercapto-Ethyl-Pyridine (MEP) resin, which shows a different salt impact trend and impurity resolution pattern from standard HIC resin, can not only capture mAb from crude CHO fermentation supernatant but also substantially enhance mAb purification process flow efficiency when serving as a polishing role.
Chen
Jie
J
Process Sciences Department, Dyax Corporation, Cambridge, MA 02139, USA. jchen@dyax.com
Tetrault
Jenifer
J
Ley
Arthur
A
D000645:Ammonium Sulfate / Q000737:chemistry; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000302:isolation & purification*; D016466:CHO Cells; D002846:Chromatography, Affinity / Q000379:methods; D002853:Chromatography, Liquid / Q000379:methods; D006224:Cricetinae; D003412:Cricetulus; D017077:Culture Media, Conditioned / Q000737:chemistry; D057927:Hydrophobic and Hydrophilic Interactions; D007141:Immunoglobulin Fc Fragments / Q000737:chemistry; D007074:Immunoglobulin G / Q000737:chemistry / Q000302:isolation & purification*; D015203:Reproducibility of Results; D012117:Resins, Synthetic / Q000737:chemistry*; D012680:Sensitivity and Specificity; D012685:Sepharose / Q000031:analogs & derivatives* / Q000737:chemistry; D012965:Sodium Chloride / Q000737:chemistry; D013205:Staphylococcal Protein A / Q000737:chemistry
D000911:Antibodies, Monoclonal; D017077:Culture Media, Conditioned; D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G; D012117:Resins, Synthetic; D013205:Staphylococcal Protein A; D012965:Sodium Chloride; C020927:phenyl-sepharose; C052754:butylagarose; D012685:Sepharose; D000645:Ammonium Sulfate