DEAE Cellulose
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Diethylaminoethyl cellulose (DEAE-C) is a positively charged
resin A resin is a solid or highly viscous liquid that can be converted into a polymer. Resins may be biological or synthetic in origin, but are typically harvested from plants. Resins are mixtures of organic compounds, predominantly terpenes. Commo ...
used in
ion-exchange chromatography Ion chromatography (or ion-exchange chromatography) is a form of chromatography that separates ions and ionizable polar molecules based on their affinity to the ion exchanger. It works on almost any kind of Charge (chemistry), charged molecule ...
, a type of
column chromatography Column chromatography in chemistry is a chromatography method used to isolate a single chemical compounds, chemical compound from a mixture. Chromatography is able to separate substances based on differential absorption of compounds to the adsorbe ...
, for the separation and purification of proteins and nucleic acids. Gel matrix beads are derivatized with
diethylaminoethanol Diethylethanolamine (DEAE) is the organic compound with the molecular formula . A colorless liquid, is used as a precursor in the production of a variety of chemical commodities such as the local anesthetic procaine. Applications Diethylethano ...
(DEAE) and lock negatively charged proteins or nucleic acids into the matrix. The proteins are released from the resin by increasing the salt concentration of the solvent or changing the pH of the solution as to change the charge on the protein.


Preparation

DEAE-C is synthesized by an alkali-catalyzed reaction of
cellulose Cellulose is an organic compound with the chemical formula, formula , a polysaccharide consisting of a linear chain of several hundred to many thousands of glycosidic bond, β(1→4) linked glucose, D-glucose units. Cellulose is an important s ...
(obtained from cotton fabric) with 2-chlorotriethylamine, illustrated as following :


Types


Common resins

DEAE-C is commonly commercially available as DE52 and DE53. These resins are prepared preswollen although cellulose exchangers swell in a strong basic environment to increase access to binding sites. DE52 has a
pKa In chemistry, an acid dissociation constant (also known as acidity constant, or acid-ionization constant; denoted ) is a quantitative measure of the strength of an acid in solution. It is the equilibrium constant for a chemical reaction :H ...
of 11.5. The buffering range for
diethanolamine Diethanolamine, often abbreviated as DEA or DEOA, is an organic compound with the formula HN(CH2CH2OH)2. Pure diethanolamine is a white solid at room temperature, but its tendencies to absorb water and to supercool often results in it being foun ...
is 8.4-8.8, though the range for DEAE-C varies between manufacturers.


DEAE-D

DEAE-
Dextran Dextran is a complex branched glucan (polysaccharide derived from the condensation of glucose), originally derived from wine. IUPAC defines dextrans as "Branched poly-α-d-glucosides of microbial origin having glycosidic bonds predominantly C-1 ...
(DEAE-D) is a positively charged dextran derivative that can be used for
vaccine A vaccine is a biological Dosage form, preparation that provides active acquired immunity to a particular infectious disease, infectious or cancer, malignant disease. The safety and effectiveness of vaccines has been widely studied and verifi ...
production,
gene therapy Gene therapy is Health technology, medical technology that aims to produce a therapeutic effect through the manipulation of gene expression or through altering the biological properties of living cells. The first attempt at modifying human DNA ...
, protein stabilization,
dyslipidemia Dyslipidemia is a metabolic disorder characterized by abnormally high or low amounts of any or all lipids (e.g. fats, triglycerides, cholesterol, phospholipids) or lipoproteins in the blood. Dyslipidemia is a risk factor for the development of ...
prevention, flocculating agents, and many other applications. DEAE-D is also used for transfecting animal cells with foreign DNA. It is added to solution containing DNA meant for transfection. It binds and interacts with negatively charged DNA molecules and via an unknown mechanism brings about the uptake of
nucleic acid Nucleic acids are large biomolecules that are crucial in all cells and viruses. They are composed of nucleotides, which are the monomer components: a pentose, 5-carbon sugar, a phosphate group and a nitrogenous base. The two main classes of nuclei ...
s by the cell. This procedure is highly suited for transient transfection used for various
molecular biology Molecular biology is a branch of biology that seeks to understand the molecule, molecular basis of biological activity in and between Cell (biology), cells, including biomolecule, biomolecular synthesis, modification, mechanisms, and interactio ...
studies.


Other derivatives

DEAE-Sepharose, DEAE-650 and DEAE-
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 ...
are commonly used in chromatography.


Ion-exchange chromatography

DEAE-C is a weak anion exchanger. This exchange is utilized to separate proteins that have faintly differing charges. Like all anion exchangers, the resin carries a positive charge that interacts favorably with negative charges. The positive charge of DEAE cellulose is due to a protonated amine group. To ensure that the resin is protonated and positively charged, the chromatography should be performed at least 2 pH units below the
pKa In chemistry, an acid dissociation constant (also known as acidity constant, or acid-ionization constant; denoted ) is a quantitative measure of the strength of an acid in solution. It is the equilibrium constant for a chemical reaction :H ...
of the amine group, 10. The strength of the bond between the resin and protein is highly dependent on the pH range in the column and the pI of the protein of interest. The resin is a weak exchanger because it is only partially ionized over most pH values, and an efficient separation with DEAE-C
chromatography In chemical analysis, chromatography is a laboratory technique for the Separation process, separation of a mixture into its components. The mixture is dissolved in a fluid solvent (gas or liquid) called the ''mobile phase'', which carries it ...
requires a specific, narrow pH range. Cellulose, dextran, agarose, and other insoluble complexes are unaffected because they compose inert matrices, hence why they are so often derivatized with strong and weak cation and anion exchangers in chromatography. DEAE-C beads have diethylaminoethyl chains covalently bound to oxygen atoms on the
D-glucose Glucose is a sugar with the molecular formula , which is often abbreviated as Glc. It is overall the most abundant monosaccharide, a subcategory of carbohydrates. It is mainly made by plants and most algae during photosynthesis from water and ...
subunits of
cellulose Cellulose is an organic compound with the chemical formula, formula , a polysaccharide consisting of a linear chain of several hundred to many thousands of glycosidic bond, β(1→4) linked glucose, D-glucose units. Cellulose is an important s ...
.


See also

*
Size-exclusion chromatography Size-exclusion chromatography, also known as molecular sieve chromatography, is a chromatography, chromatographic method in which molecules in Solution (chemistry), solution are separated by their Chemical structure, shape, and in some cases molec ...
* Stationary phase


References

{{reflist Cellulose ethers