ABSTRACT

Size exclusion chromatography (SEC) is a mode of liquid chromatography in which separation is based on the partial exclusion of the solutes from the pores of the stationary phase [1]. The larger molecules elute earlier than the smaller ones, because large solutes can penetrate into the pores only by a smaller fraction (partially) [1]. Very large solutes (larger than the pores) can be totally excluded from the pores and will elute within the interstitial column volume. Based on its mechanism, SEC is applied to separate solutes of different sizes (molecular weights). Hence, SEC is commonly used to separate protein aggregates (size variants) such as monomers, dimers, trimers, and other oligomers or high-molecular-weight species (HMWS).