Size Exclusion Chromatography, Principles and Methods

This handbook describes the use of size exclusion chromatography (SEC) for the purification and separation of biomolecules, with focus on practical information for optimized results. SEC, which is also known as gel filtration chromatography, separates biomolecules by differences in size as they pass through a resin packed in a chromatography column. SEC differs from techniques such as as ion exchange (IEX) or affinity chromatography (AC) in that molecules do not bind to the chromatography resin. The technique is an excellent choice for high-resolution separation of biomolecules with low consumption of buffer. Analytical SEC is used for high-resolution purification of biomolecules, rapid purity checks, and screening of proteins. SEC may also be used for group separation of smaller biomolecules such as salts from larger biomolecules such proteins in desalting/buffer exchange.