Sephadex column chromatography

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],... 

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). 

Sephadex column chromatography of dansvlated glvconep-tide surfactant... 

Bleomycin was isolated from the culture filtrate of Streptomyces verticillus by ion exchange resin adsorption, alumina and Sephadex column chromatography. Bleomycin thus obtained was a mixture of more than 10 components, which can be separated on a CM-Sephadex column by elution with a linear gradient of ammonium formate5) (see Fig. 1). Later, it was found that they are different from one another in... 

Fig. 6. Acrylamide gel electrophoresis of glycoproteins separated by QAE-Sephadex column chromatography shown in Fig. 5. Strips of Whatman No. 3 filter paper, 1 mm X 1 cm, were dipped into 5% solutions of the glycoproteins and inserted into slots at the origin. The gel was stained for carbohydrate with a-naphthol and concentrated HjS04. The unlabeled fraction shows glycoproteins 1-5, which were chromatographed on the QAE-Sephadex column. Fraction III from Sephadex contained glycoprotein 3, fraction V contained glycoprotein 4, and fraction VII contained glycoprotein 5. From DeVries et al. (1970), reproduced with permission.

In a separate study, the quantity of thalidasine in a polyphase-liposome injection (139-2) was determined by dilution of the injection with EtOH-O.OlN HC1 (4 1), followed by spectrofluorometric determination at 235 nm (for excitation) and at 318 nm (for emission). The recovery was > 96% and the coefficient of variation was < 3%. The polyphase-liposome encapsulation rate was also determined by this method after separation via Sephadex column chromatography [148]. 

Resolution (or separation) of optical isomers was achieved by SP-Sephadex column chromatography using 0.02M Na2[Sb2(L-tart)2] For the N-C-C-P and the P-C-C-P complexes, the bands of optical isomers were separated incompletely, so that the chromatography was repeated until no further increase in Ae/e values was observed. From the eluates, the optical isomers were isolated by a method similar to that for the racemates. 

In the [Co(en)j(N—N)] " complexes, two nitrogen atoms of the N—N ran occupy either cis or tram position as shown in Fig. 1.5 a, if the methylenic chain is long enough. However, from the fact that the [Co(en)2(N—N)] " complexes (N—N = tmd and don) were resolvte into optical isomers by SP-Sephadex column chromatography the workers assigned cis structure to all the complexes. For the dimeric complexes, [Co2(ai)b(N—structures shown in Fig. 1.5 b were proposed as most probable on the basis of molecular models. 

Amylose of DP ca. 100 and amylopectin were corn products purchased from Kanto Chemical Co., Inc. Other kinds of amylose from corn were obtained from Hayashibara Biochemical Laboratories and from Dr. John Robyt of Iowa State University, U.S.A. These samples of amylose were separated using the Sephadex column chromatography and were certified to have narrow distribution of molecular weight. Sodium hydroxide and other chemicals were guaranteed products of Wako Pure Chemical Industries Ltd. 

Isolation of individual iridoids by traditional silica gel/alumina/charcoal/ dianion/Sephadex column chromatography is tedious because of their close Rf values and polar nature. Application of HPLC techniques using octadecyl-silanized (0DS)Si02/CigSi gel colunm or HPLC coupled with a diode-array detector and a mass spectrometer (HPLC-DAD-MS) has achieved efficient separation... 

The brown solution from the fifth band obtained by the first Sephadex column chromatography turns blue-violet within several hours, and the resulting blue-violet solution containing W3 can be purified similarly to yield ca. 5% W3. 

Musa et al. (1965) reported the purification of human liver jS-glucuroni-dase using DEAE phadex and CM-Sephadex column chromatography. The final product, a single protein, had a specific activity of 57,500 and was characterized. 

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