Enzyme Purification and Concentration by Simulated Moving Bed Chromatography: An Experimental Study

Abstract

A new continuous affinity chromatography for enzyme purification and concentration was proposed. It combined the features of pH-parametric pumping and simulated moving bed operations. Adsorption and desorption were taking place simultaneously in the system. Regeneration of adsorbent was performed by successively shifting the adsorption and desorption sections from one column to another with a time controller. Performance tests for enzyme separation and purification were carried out by devising six-staged affinity chromatographic columns, 16cm ID x 40cm Ht, packed with lOcm height chitin-CHOM each for trypsin purification. Factors affecting the successful operations of the systems and separation efficiency of enzyme were investigated. The system was applied to the purification of trypsin from extract of porcine pancreas. It was found that fairly high separation efficiency can be achieved, SAR (specific activity ratio = specific activity of productlspecific activity of feed) of eluant was as high as 3.9. Thus it is considered as a promising method of enzyme purification. For purification of c~d e enzyme, SMB (Simulated Moving Bed) can be incorporated with a washing section between the adsorption and desorption sections for washing out the impure proteins contaminated into the columns.

Continuous annular chromatography: General characterization and application for the isolation of recombinant protein drugs

“…Several continuous separation technologies like simulated moving bed chromatography (Juza et al, 2000;Schulte and Strube, 2001;Nicoud, 2000;Jensen et al, 2000;Gottschlich et al, 1996;Hashimoto et al, 1988;Huang et al, 1986), continuous extractive techniques (Jarudilokkul and Stuckey, 2001;Carneiro-da-Cunha et al, 1996;Pires et al, 1996;Asenjo, 1994;Veide et al, 1984), as well as continuous annular chromatography (Fox et al, 1969;Bloomingburg et al, 1991;Bloomingburg and Carta, 1994) can in principle all be applied or adapted for isolation of recombinant proteins. More recently, continuous annular chromatography (CAC) became commercially available (P-CAC, preparative continuous annular chromatography; Prior Separation Technology, Austria).…”

Experimental evaluation of designs for the simulated countercurrent moving bed separator

“…Small-scale continuous simulated countercurrent processes have been described and are operated in a similar fashion to the Sorbex separators (Broughton, 1984;Ching et al, 1985~;De Rosset et al, 1981;Huang et al, 1986;Kubota et al, 1989;Ruthven, 1984;Ruthven and Ching, 1989;Saska et al, 1992;Storti et al, 1992). Either a single long column with multiple feed introduction ports, or multiple columns and multiple feed ports are used in these apparatuses.…”