Chromatography Columns and Media

In column chromatography, the stationary bed that defines the separation chemistry is generally a tube or a column. The appropriate type of column and separation media will depend on the size, charge, affinity, mass, and other physical properties of the analytes. Empty columns can be packed with a media of choice by skilled technicians; busy laboratories can use pre-packaged reusable and disposable columns to increase throughput. Low-pressure spin columns can be run in a benchtop centrifuge; larger columns can be run under gravity, vacuum, or a medium to high-pressure chromatography system. Pre-packed columns are available for all separation methods such as affinity purification, size exclusion, reverse phase, ion exchange, and hydrophobic interaction. Important considerations when choosing a column are the desired final product purity, the physical properties of the protein of interest, the chromatography system in use, and the budget for the separation process.