Hydrophobic interaction chromatography preparative

The cytokine-inducing active fraction (QM-A) was prepared from E, hirae ATCC 9790 cells as described [9]. Briefly, the bacterial cells (922 g) were delipidated and extracted with hot phenol-water, and the crude extract was digested with RNase and DNase, to give crude LTA fractions. The crude LTA was subjected to two successive chromatography, hydrophobic interaction... 

The chemical adsorption of a relatively high molecular weight neutral polymer (poly(succinimide), M = 13000) on aminopropyl-Vydac 101 TP silica gel was applied by Alpert [47, 48] to prepare a reactive composite support for use in cation-exchange [47] and hydrophobic-interaction [48] chromatography of pro-... 

Fig. 8. Preparative isolation of hexon antigen of EDS-76 by hydrophobic-interaction chromatography on Butyl-PG column (2x5 cm) (A) application of the allantoic fluid diluted (1 5) by 50 mM potassium acetate, pH 4,130 ml (B)0.01 mol/1 potassium acetate, pH 5.5 (C) 0.01 mol/1 potassium bicarbonate pH 8.0, 10% isopropanol (D) 0.01 mol/1 potassium carbonate pH 9.6, 10% isopropanol. EDS-0 — components of alantoic fluid eluted with buffer A, EDS-1 — desorbed hexon fraction eluted with buffer C, EDS-2 — fraction desorbed with buffer D [56]...

After initial cell fermentation and product extraction from the producer cells, the crude preparation is subject to multiple chromatographic steps, including ion-exchange, hydrophobic interaction chromatography and gel-filtration chromatography. The purified product is presented in lyophilized form in vials (1 mg active/vial) and excipients include a phosphate buffer, sodium chloride and serum albumin. 

Table 4. Purification of (S)-ADH from Rhodococcus erythropolis DSM 43 297 (1 U = reduction of 1 pmol p-Cl-acetophenone per min HIC = hydrophobic interaction chromatography Prep-Cell = preparative gel electrophoresis)...

Miller, N.T. and Shieh, C.H., Preparative hydrophobic interaction chromatography of proteins using ether based chemically bonded phases, J. Liq. Chromatogr., 9, 3269, 1986. 

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