pH–conductivity hybrid gradient cation-exchange chromatography for process-scale monoclonal antibody purification

Recovery and purification process development for monoclonal antibody production

“…Generally, a linear salt or pH gradient elution program can be conducted to determine the best elution condition. [55][56][57][58] An elution condition development study conducted for an IgG1 is shown in Figure 6. Four runs were performed in this study with two Fractogel cation exchange resins, Fractogel COO -(weak cation exchanger) chromatographic step such as cation exchange, so it may not be applicable to all products.…”

Adsorption equilibrium and kinetics of monomer–dimer monoclonal antibody mixtures on a cation exchange resin

“…Ansaldi and Lester [9], for example, have described the use of salt steps and salt gradients to separate dimers and multimers from protein monomers on both weak and strong CEX resins. Zhou et al [10] demonstrated aggregate clearance from a mAb feedstock utilizing salt gradients and hybrid pH-salt gradients with both weak and http://dx.doi.org/10.1016/j.chroma.2015.05.007 0021-9673/© 2015 Elsevier B.V. All rights reserved. strong CEX resins.…”

Ion Exchange Chromatography of Proteins and Clearance of Aggregates

“…Elution is achieved with either increasing linear salt gradient or step elution with predetermined salt concentration. In certain instances elution can be achieved with increasing elution buffer pH [12]. CEX can also be operated in flow-through mode, were the load is conditioned at a pH and salt concentration which would allow the product to flowthrough while the impurities are retained on the column.…”

“…Alternatively the protein could be eluted with an increasing pH gradient [12]. Increased elution buffer pH weakens the interaction between the protein and the resin and offers an added benefit of reducing the elution salt concentration.…”