Sepharose is a tradename for a crosslinked, beaded-form of
agarose
Agarose is a heteropolysaccharide, generally extracted from certain red algae. It is a linear polymer made up of the repeating unit of agarobiose, which is a disaccharide made up of D-galactose and 3,6-anhydro-L-galactopyranose. Agarose is on ...
, a polysaccharide polymer material extracted from seaweed. Its brand name is a
portmanteau
In linguistics, a blend—also known as a blend word, lexical blend, or portmanteau—is a word formed by combining the meanings, and parts of the sounds, of two or more words together. derived from Separation-Pharmacia-Agarose. A common application for the material is in chromatographic separations of biomolecules.
Sepharose is a registered trademark of
Cytiva
Danaher Corporation is an American global conglomerate founded in 1984 by brothers Steven and Mitchell Rales. Headquartered in Washington, D.C., the company designs, manufactures, and markets medical, industrial, and commercial products and ...
(formerly:
GE Healthcare and
Pharmacia
Pharmacia was a pharmaceutical and biotechnological company in Sweden that merged with the American pharmaceutical company Upjohn in 1995.
History
Pharmacia company was founded in 1911 in Stockholm, Sweden by pharmacist Gustav Felix Grönfel ...
, Pharmacia LKB Biotechnology, Pharmacia Biotech, Amersham Pharmacia Biotech, and Amersham Biosciences).
Various grades and chemistries of sepharose are available.
Iodoacetyl functional groups can be added to selectively bind
cysteine
Cysteine (; symbol Cys or C) is a semiessential proteinogenic amino acid with the chemical formula, formula . The thiol side chain in cysteine enables the formation of Disulfide, disulfide bonds, and often participates in enzymatic reactions as ...
side chains and this method is often used to immobilize
peptide
Peptides are short chains of amino acids linked by peptide bonds. A polypeptide is a longer, continuous, unbranched peptide chain. Polypeptides that have a molecular mass of 10,000 Da or more are called proteins. Chains of fewer than twenty am ...
s. Sepharose/agarose, combined with some form of activation chemistry, is also used to immobilize
enzyme
An enzyme () is a protein that acts as a biological catalyst by accelerating chemical reactions. The molecules upon which enzymes may act are called substrate (chemistry), substrates, and the enzyme converts the substrates into different mol ...
s,
antibodies
An antibody (Ab) or immunoglobulin (Ig) is a large, Y-shaped protein belonging to the immunoglobulin superfamily which is used by the immune system to identify and neutralize antigens such as bacteria and viruses, including those that caus ...
and other
proteins
Proteins are large biomolecules and macromolecules that comprise one or more long chains of amino acid residues. Proteins perform a vast array of functions within organisms, including catalysing metabolic reactions, DNA replication, re ...
and peptides through
covalent
A covalent bond is a chemical bond that involves the sharing of electrons to form electron pairs between atoms. These electron pairs are known as shared pairs or bonding pairs. The stable balance of attractive and repulsive forces between atom ...
attachment to the resin. Common activation chemistries include
cyanogen bromide
Cyanogen bromide is the inorganic compound with the chemical formula, formula BrCN. It is a colorless solid that is widely used to modify biopolymers, fragment proteins and peptides (cuts the C-terminus of methionine), and synthesize other compo ...
(CNBr) activation and reductive amination of
aldehydes
In organic chemistry, an aldehyde () (lat. ''al''cohol ''dehyd''rogenatum, dehydrogenated alcohol) is an organic compound containing a functional group with the structure . The functional group itself (without the "R" side chain) can be referred ...
to attach proteins to the agarose resin through lysine side chains.
See also
*
Gel permeation chromatography
Gel permeation chromatography (GPC) is a type of size-exclusion chromatography (SEC), that separates high molecular weight or colloidal analytes on the basis of size or diameter, typically in organic solvents. The technique is often used for the an ...
*
Superose Superose is a trade name for a collection of FPLC columns which are used in the automated separation of biological molecules. The different columns provided can separate a variety of macromolecules, ranging from small peptides and polysaccharides t ...
*
Sephadex Sephadex is a cross-linked dextran gel used for gel filtration. It was launched by Pharmacia
Pharmacia was a pharmaceutical and biotechnological company in Sweden that merged with the American pharmaceutical company Upjohn in 1995.
History ...
References
Polysaccharides
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