Sephadex is a cross-linked dextran gel used for gel filtration. It was launched by Pharmacia in 1959, after development work by Jerker Porath and Per Flodin.[1][2] The name is derived from separation Pharmacia dextran. It is normally manufactured in a bead form and most commonly used for gel filtration columns. By varying the degree of cross-linking, the fractionation properties of the gel can be altered.
These highly specialized gel filtration and chromatographic media are composed of macroscopic beads synthetically derived from the polysaccharide dextran. The organic chains are cross-linked to give a three-dimensional network having functional ionic groups attached by ether linkages to glucose units of the polysaccharide chains.
Available forms include anion and cation exchangers, as well as gel filtration resins, with varying degrees of porosity; bead sizes fall in discrete ranges between 20 and 300 μm.
Sephadex is also used for ion-exchange chromatography.[3]
Sephadex is crosslinked with epichlorohydrin.[4]
Applications
editSephadex is used to separate molecules by molecular weight. Sephadex is a faster alternative to dialysis (de-salting), requiring a low dilution factor (as little as 1.4:1), with high activity recoveries. Sephadex is also used for buffer exchange and the removal of small molecules during the preparation of large biomolecules, such as ampholytes, detergents, radioactive or fluorescent labels, and phenol (during DNA purification).
A special hydroxypropylated[5] form of Sephadex resin, named Sephadex LH-20, is used for the separation and purification of small organic molecules such as steroids, terpenoids, lipids. An example of use is the purification of cholesterol.[6]
Fractionation
editExclusion chromatography.
Fractionation Range[7] of Globular Proteins and Dextrans (Da).
| Gel Type | Fractionation Range | |
|---|---|---|
| Globular
Proteins |
Dextrans | |
| G-10 | ≤700 | ≤700 |
| G-15 | ≤1500 | ≤1500 |
| G-25 | 1000–5000 | 100–5,000 |
| G-50 | 1500–30,000 | 500–10,000 |
| G-75 | 3000–80,000 | 1000–50,000 |
| G-75 SF | 3000–70,000 | 1000–50,000 |
| G-100 | 4000–150,000 | 1000–100,000 |
| G-100 SF | 4000–100,000 | 1000–100,000 |
| G-150 | 5000–300,000 | 1000–150,000 |
| G-150 SF | 5000–150,000 | 1000–150,000 |
| G-200 | 5000–600,000 | 1000–200,000 |
| G-200 SF | 5000–250,000 | 1000–200,000 |
Ion-exchange chromatography.
| Description | Functionality | Bead size (μ) |
|---|---|---|
| Sephadex-CM C-25 | carboxymethyl | 40-120 |
| Sephadex-CM C-50 | carboxymethyl | 40-120 |
| Sephadex-DEAE A-25 | 2-(diethylamino)ethyl | 40-120 |
| Sephadex-DEAE A-50 | 2-(diethylamino)ethyl | 40-120 |
| Sephadex-QAE A-25 | quaternary aminoethyl | 40-120 |
| Sephadex-QAE A-50 | quaternary aminoethyl | 40-120 |
| Sephadex-SP C-25 | sulfopropyl | 40-120 |
| Sephadex-SP C-50 | sulfopropyl | 40-120 |
See also
editReferences
edit- ^ "From Sephadex to GE Healthcare". chemeurope.com. Archived from the original on July 31, 2012. Retrieved Jan 2, 2019.
- ^ Porath, J; Flodin, P (1959). "Gel Filtration: A Method for Desalting and Group Separation". Nature. 183 (4676): 1657–1659. Bibcode:1959Natur.183.1657P. doi:10.1038/1831657a0. ISSN 1476-4687. PMID 13666849. S2CID 32287460.
- ^ "Sephadex® ion exchange media" (PDF). chembio.uoguelph.ca. Pharmacia Biotech.
- ^ "GE Healthcare Life Sciences - Instructions for Sephadex Media". .gelifesciences.com. Archived from the original on 2012-02-18. Retrieved 2011-12-02.
- ^ GE Healthcare. "Sephadex LH-20". Retrieved 2019-12-06.
- ^ Hanukoglu I, Jefcoate CR (1980). "Pregnenolone separation from cholesterol using Sephadex LH-20 mini-columns". Journal of Chromatography A. 190 (1): 256–262. doi:10.1016/S0021-9673(00)85545-4. ISSN 0021-9673.
- ^ "Instructions for Sephadex Media" (PDF). gelifesciences.co.jp (product manual). GE Healthcare. Apr 7, 2008. Retrieved Jan 2, 2019.