Size-Exclusion Chromatography

Size-exclusion chromatography (also known as gel filtration chromatography) is a technique for separating proteins and other biological macromolecules on the basis of molecular size. Originally developed in the 1950s, the technique was developed using crosslinked dextran ( 1 , 2 ). Some confusion over nomenclature has been created by the term gel permeation, used to describe separation by the same principle in organic mobile phases using synthetic matrices ( 3 ). It is now generally agreed that the terms gel filtration and gel permeation do not accurately reflect the nature of the separation. Size-exclusion chromatography (SEC) has been widely accepted as a universal description of the technique and in line with the IUPAC nomenclature, this term will be adopted ( 4 ). Size-exclusion chromatography is a commonly used technique, because of the diversity of the molecular weights of proteins in biological tissues and extracts. SEC has been employed for many roles, including buffer exchange (desalting), removal of nonprotein contaminants (DNA, viruses) ( 5 ), protein aggregate removal ( 6 ), the study of biological interactions ( 7 – 9 ), and protein folding ( 10 – 12 ). It also has the important advantage of being compatible with physiological conditions ( 13 , 14 ). These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.