Fundamentals Of Size Exclusion Chromatography

Size exclusion chromatography (SEC), also known as gel filtration, is the mildest of all the chromatography techniques.

Molecules are separated by size differences as they pass through a resin packed in a column, and molecules do not bind to the chromatography resin, meaning buffer composition does not directly affect resolution, which is the degree of separation between peaks. This differs from techniques such as ion exchange chromatography (IEX) or affinity chromatography (AC). Consequently, conditions can be varied to suit the type of sample or the requirements for further purification, analysis, or storage without altering the separation.

You can use SEC for both preparative and analytical studies of your protein. But how do you know which parameters will give you the best resolution of your protein mixture?