A microscale approach for predicting the performance of chromatography columns used to recover therapeutic polyclonal antibodies

A microscale approach is described which screens conditions for recovering polyclonal antibodies from ovine sera by mixed-mode cation-exchange chromatography. The impact of pH and loading buffer salt concentration were assessed using robotically operated 20 μL packed pipette tips. Low salt concentrations delivered capacities up to 41 mg/mL, while only half this level was obtained at high salt concentrations. Two of the screened conditions were then tested in a 10 mL packed bed and overall trends in capacity, yield and purity were found to be retained. Microscale pipette tips thus provided a useful basis for the rapid, approximate definition of a chromatography design space.