Optimization of monoclonal antibody purification by ion-exchange chromatography. Application of simple methods with linear gradient elution experimental data.
Simple methods for the optimization of ion-exchange chromatography of proteins in our previous papers were applied to cation-exchange chromatography purification of monoclonal antibodies (Mab). We carried out linear gradient elution experiments, and obtained the data for the peak salt concentration and the peak width. From these data, the distribution coefficient as a function of salt concentration, and the height equivalent to a theoretical plate (HETP) as a function of mobile phase velocity were calculated. The optimized linear gradient elution conditions were determined based on the relationship between buffer consumption and separation time. The optimal stepwise elution conditions were determined based on the relationship between the distribution coefficient and the salt concentration.
Ishihara
Takashi
T
Kirin Brewery Co. Ltd., Takasaki, Gunma 370-0013, Japan. t-ishihara@kirin.co.jp
Yamamotob
Shuichi
S
D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D002411:Cation Exchange Resins; D002852:Chromatography, Ion Exchange / Q000379:methods*; D008958:Models, Molecular
D000911:Antibodies, Monoclonal; D002411:Cation Exchange Resins