In gel filtration chromatography, the stationary phase is comprised of porous beads packed into a column. The mobile phase is the running buffer or other solvent. Sample components partition between the stationary and mobile phases based on their size-based accessibility to the pores of the matrix beads.
Size Exclusion Chromatography (Gel Filtration)
Size exclusion chromatography (SEC) separates components of a sample on the basis of their molecular size. Differential exclusion or inclusion of the molecules is achieved via filtration through a gel that contains spherical beads. These beads have pores of a specific size distribution so as to include or exclude molecules of different sizes when they pass through the gel.
Learn More in these related Britannica articles:
Size Exclusion Chromatography (SEC) is a chromatographic method which separates analytes solely based on their size, where molecules are separated on the basis of their exclusion from pores in the column packing material. Larger analytes will elute first, while the smaller molecules interact more with the stationary phase and will elute later.
Structural Biochemistry-Proteins-Purification-Gel-Filtration chromatography
Many size exclusion chromatography columns have limitations on the eluents that can be used. Using an eluent that is not specifically permitted could irreversibly expand the packing material or otherwise cause physical deterioration. Therefore, investigate which solvents readily dissolve the sample in advance and then choose a column compatible with one of those solvents.
Protein Purification Methods
In a group separation, the sample components are separated into two major groups according to size range. Group separation can be used to remove high-or low-molecular weight impurities (such as for example phenol red from culture fluids) or for desalting and buffer exchange.
Gel Filtration Chromatography
Gel filtration, on the other hand, uses size exclusion to separate molecules of different dimensions. The range of charged porous particles in the gel filtration media determines the size of molecules that will be separated. Molecules that are smaller than the average or maximum effective pore size (also known as the fractionation range or exclusion limit of the resin) permeate through the pores of the gel filtration media wherein they follow a complex path that slows down their migration through the resin bed.
People also read
Gel filteration chromatography is also known as Gel permiation chromatography or size exclusion chromatography. Unlike SDS-PAGE which separates the denatured protein based on mass, size exclusion chromatography separates the protein molecules based on its mass and shape. It uses small polymeric beads made up agarose (Sepharose), Dextran (Sephadex), polyacrylamide, superdex, etc. These beads have fixed pore size. The pore size is such that the molecules which are smaller or nearly equal can enter.
Gel Filtration Chromatography Technique as Tool of Simple Study Seminal Plasma Proteins in Domestic Animals
Citation: Belico de VA, Silvano de OJ, de Albuquerque LM (2015) Gel Filtration Chromatography Technique as Tool of Simple Study Seminal Plasma Proteins in Domestic Animals. J Chromatogr Sep Tech 6:281. doi:10.4172-2157- 7064.1000281