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Column chromatography

Column chromatography separates molecules in a liquid sample from each other by applying a mobile phase onto the stationary-phase column. The technique generally separates compounds based on differential adsorption to the ligand on the chromatography resin. The exception is size exclusion chromatography, where molecules don’t interact with the ligand on the column; instead, they are separated based on size. Column chromatography is used for analytical purposes and lab-scale purification in life sciences labs. It’s also used in biomanufacturing for process development – for both analysis and purifcation - and for large-scale purification.

Empty chromatography columns FAQs

Here are answers to some frequently asked questions.

What are empty chromatography columns?

Empty chromatography columns for lab scale are glass columns used during liquid chromatography to separate molecules such as proteins from other molecules and substances such as salts. Process-scale empty columns are also available for large-scale purification. Column packing is required prior to use manually or with a liquid chromatography system. The chromatography resin of choice – perhaps an ion exchange resin or hydrophobic interaction resin – is added to the empty column in a controlled and consistent way.

What are the basic principles of column chromatography?

In column chromatography, solutes in the applied sample interact with the ligand of the chromatography resin inside the column. After sample loading, the column is washed with a mobile phase to remove molecules that don’t interact strongly. Then another mobile phase, the elution buffer, provides conditions to desorb the remaining molecules from the ligand.

What are the applications of chromatography columns?

Applications of empty chromatography columns vary widely in the pharmaceutical and petroleum industries, environmental or life sciences, and toxicology. The most common uses of column chromatography in life sciences and the biopharma industry are:

  • Purification of molecules, for example, protein purification of antibodies
  • Analysis of a sample to determine purity level
What goes into a chromatography column?

Liquid chromatography often involves the use of small particles or resins, which get packed and held in a column; this is also referred to as the stationary phase. Resins are based on different materials, such as agarose or dextran, which are prepared to have the desired physical and chemical characteristics.

What types of resins can be used?

Based on chromatography techniques, there is a diverse array of resins or media available to help purify a broad range of target molecules. Common types of chromatography resins to capture biomolecules include:

What are chromatography columns made of?

Empty chromatography columns often comprise glass or stainless-steel materials. Some fast protein liquid chromatography columns come in polypropylene. You can choose to pack your own columns or purchase prepacked columns that are ready for use.

How to choose an empty column

Use the Purify app to help you find the right Cytiva column for your application.