ABSTRACT
Size exclusion chromatography 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. The larger molecules elute earlier than the smaller ones, because large solutes can penetrate into the pores only by a smaller fraction. The stationary phase consists of spherical porous particles with a carefully controlled pore size and pore size distribution, through which the solutes diffuse based on their molecular size differences, using an aqueous buffer as the mobile phase. Entropic processes without any physical or physicochemical adsorption ideally drive the partitioning between the internal pores and interstitial volume. Resolution between the aggregates and native protein peaks can be improved by decreasing the column particle size. The achievable plate number is indeed inversely proportional to the particle size, while the resolution is inversely proportional to the square root of the particle size.
