DEAE-Sephadex column

Coiicin E (from E.coli) [11032-88-5], Purified by salt extraction of extracellular-bound colicin followed by salt fractionation and ion-exchange chromatography on a DEAE-Sephadex column, and then by CM-Sephadex column chromatography [Schwartz and Helinski J Biol Chem 246 6318 1971],... 

Reverse transcriptase (from avian or murine RNA tumour viruses) [9068-38-6] [EC 2.7.7.49]. Purified by solubilising the virus with non-ionic detergent. Lysed virions were adsorbed on DEAE-cellulose or DEAE-Sephadex columns and the enzyme eluted with a salt gradient, then chromatographed on a phosphocellulose column and enzyme activity eluted in a salt gradient. Purified from other viral proteins by affinity chromatography on a pyran-Sepharose column. [Verna Biochim Biophys Acta 473 1 7977 Smith Methods Enzymol 65 560 1980 see commercial catalogues for other transcriptases.]... 

Chromatography of hemoglobins on columns of DEAE-Cellulose (DE-52 mlcrogranular, preswollen Whatman) often results In excellent separations of many variants, because the hemoglobins are eluted as sharp, narrow zones generally widely separated from each other The hemoglobin zones are eluted from the DE-52 columns at a distinctly higher pH value than from a similar DEAE-Sephadex column ... 

Figure 6. Isoelectric focusing of the Ci component of F. solani cellulase in an ampholyte solution covering the pH range 4.6-5.1 in an LKB 110-mL column. The C. component used was that which was eluted from a DEAE-Sephadex column as a single component (12). Activity to HsP04-swollen cellulose (- A activity to Avicel with CL and /3-glucosidase added to show synergistic activity (- O -) pH gradient (- -). From...

Application of buffered tetrahydrofuran extraction to gastric mucosa, followed by careful examination of the aqueous phase for various glycosphingolipids, indicated that this phase contained sialoglycosphingolipids and considerable quantities of neutral glycosphingolipids (22). These were separated from the acidic glycosphingolipids by DEAE-Sephadex column chromatography (23). 

Fig. 5.3. Dependence of elution position on charge. Oligonucleotides in 7 M urea on DEAE-Sephadex columns at pH 2.7.

Ion Exchange on DEAE Sephadex Columns. The use of various types of DEAE Sephadex ion-exchangers has been reported lately. Pettersson (36) found DEAE Sephadex A-25 to be a convenient material for the... 

Figure 2 (A) Eiution profiie of 6-thiopurines obtained on a iow-pressure DEAE-Sephadex column showing times of 90 fractions coiiected. (B) Liquid chromatograms of seven coiiected fractions showing content of specific 6-thiopurines. GMP, guanosine-5 -monophosphate GDP, guanosine-5 -diphosphate GTP, guanosine-5 -triphosphate. (Reproduced with permission from Breter H and Mertes H (1990) Biochimica et Biophysica Acta 1033 124-132 American Society of Health-System Pharmacists.)...

Since botulinum is a neurotoxin, large-scale production should meet biosafety level 3 containment and strict handling to avoid the possible human intoxication. Fermentation is carried out in complex medium containing casein hydrolysate, yeast extract, and glucose for 4 days. Then, botulinum toxin is precipitated with 3 N sulfuric acid followed by purification with DEAE-Sephadex column. Botulinum toxin was crystallized then and purified by additional steps using various affinity and size exclusion chromatography. Purity and quality were checked by animal testing. Production and purification steps vary with Clostridium strain used and botulinum type to be produced. ... 

Since the anion-exchange resins are frequently used in the hydroxy form, certain precautions should be observed when employing them with non-aqueous solvents. If acetone is used, excessive diacetone alcohol may be formed via base-catalyzed aldol condensation. A small amount of water should be present in all organic solvents containing alcohol to repress transesterification of esters (15) and esterification of acids (16) with alcohol as solvent. Without water, up to 5% of transesterification in 15 to 20 min has been observed with DEAE-Sephadex. After an overnight contact the transesterification approached 35 %. With 1 % water in the solvent, however, less than 0.3% transesterification and 0.3% saponification occurred when 70 mg of butyl stearate was passed through a DEAE-Sephadex column (15). 

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