| Abstract | dc.description.abstract | The behavior of ion-exchange chromatography
is well understood with respect to changes in ionic strength,
pH, resin ligand density, bed height, elution flow rate, and
gradient slope. Their relative importance for any specific
chromatographic situation varies. When a chromatographic
operation utilized to purify a human therapeutic protein is
prepared for validation before commercial production,
numerous tests have to be performed to establish the relative
importance of each operating parameter to define its future
role and importance in the framework of in-process controls.
This prioritization process is usually performed using
a purely empirical approach. In this work, we demonstrate
the application of a rational approach based on chromatographic
theory to prioritize operating parameters. Both
methodologies, empirical and rational, were performed to
evaluate a specific ion-exchange chromatography operation
for the preparative separation of closely related protein
species. We show that the application of the rational
approach has the potential to accelerate the evaluation
and significantly reduce the amount of analytical testing
needed. | en |
