Why and How to Do It

P reparative chromatography is the dominant purification technique in the production of biological compounds, especially for bioseparations. The column used is central to the performance of the chromatographic step. To ensure robust and reproducible column performance, columns need to be qualified in an acceptable fashion before each use. Some of the critical elements of column qualification include identifying leachables and extractables from resins, product use, and column parts and accessories (such as residuals from manufacturing and shipping). Identity tests must be developed for chromatography resins, such as base matrices (FTIR spectroscopy, for example); functional groups and substitution levels (such as colorimetric titration); and porosity tests, or if necessary, function tests (such as dynamic capacity, selectivity, or real runs). Column qualification also involves writing protocols for training plant personnel and protocols for column hardware, packing, and operation. And qualification requires protocols for measuring bed integrity, such as background buffers, tracers, and injection Qualification of a Chromatographic Column Why and How to Do It