Expanded-bed chromatography

Mammalian cells were cultivated in high cell density perfusion operation mode for the production of therapeutically important proteins. Material and energy balances were determined and cell regulation was investigated. Methods were developed for the recovery and purification of the products with chromatography and expanded bed chromatography. 

Vergnault, H., Mercier-Bonin, M., and Willemont, R.-M. (2004). Physicochemical parameters involved in the interaction of Saccharomyces cereviae cells with ion-exchange adsorbents in expanded bed-chromatography. Biotechnol. Prog. 20,1534-1542. 

Mattiasson, B., ed. Bioseparation International Journal of Separation Science. Special Issue Expanded Bed Chromatography. 8(1/5), The Netherlands Kluwer (1999). 

Anspach FB, Curbelo D, Hartmann R, Garke G, Deckwer WD (1999), Expanded-bed chromatography in primary protein purification, J. Chromatogr. A 865 129-144. 

There are a number of special features-challenges using expanded-bed chromatography as compared to conventional packed-bed operation. These are discussed individually in the subsequent pages, and then toward the end, some applications are presented. 

An important feature when operating expanded-bed chromatography is that the liquid is evenly distributed over the whole cross section of the column. To achieve this, certain arrangements are made. The design of proper flow distributors is an essential task. At the top it is less demanding, but one must still be secure that no mixing takes place. It is important to keep the column in a vertical mode, otherwise band-broadening may appear.4... 

The most common support for expanded-bed chromatography so far is Streamline. The material consists of crosslinked agarose with entrapped quartz crystals. The experiences of cleaning processes are clear Streamline stands up very well to the CIP treatment. In addition, ion-exchange and... 

Expanded bed chromatography for removal of cell debris with concomitant capture of target molecule. 

TABLE 2 Applications Where Expanded-Bed Chromatography has Played an Important Role... 

Expanded-bed chromatography is a young technique, but it has already proven itself a powerful and versatile tool in downstream processing work. It is likely that the technology will develop rapidly in the next few years. 

R. Semifluidized or expanded bed. Liquid-solid transfer. Na - T - 2 +1,5 (1 - eL)N Ng fljh, = pN = p/pD [E] l = liquid-phase void fraction, pp = particle density, p = fluid density, dp = particle diameter. Fits expanded bed chromatography in viscous liquids. [64] [159]... 

O. Bertrand, S. Cochet, and J. P. Cartron, Expanded bed chromatography for one-step purification of mannose binding lectin from tuhp bulbs using mannose im-mobihzed on DEAE Streamline, J. Chromatogr. A 822 19 (1998). 

Zhi9UO Su, Ph.D. Professor and Director, State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, CAS, Beijing, China (Selection of Biochemical Separation Processes—Protein Refolding, Expanded-Bed Chromatography)... 

Cross-linked cellulose based adsorbent beads have been under development at University Department of Chemical Technology (UDCT), Mumbai, and IIT, Delhi for use in expanded bed chromatography. Properties of the beads are claimed to be equivalent or even superior to the imported products. Purification protocols for expanded bed chromatography using cellulose beads for several enzymes are being developed. 

Bai Y, Glatz CE (2003) Bioprocess considerations for expanded-bed chromatography of cmde canola extract sample preparation and adsorbent reuse. Biotechnol Bioeng 81 775-782. 

Removal of cell debris Centrifugation Filtration, centrifugation or expanded bed chromatography... 

Galaev 1. Y., New methods of protein purification. Expanded bed chromatography, Biochemistry-Moscow, 63, 619-624, 1998. 

Kampen I. Einfuss der Zellaufschlussmethode auf die Expanded Bed Chromatographie. Fakultdt fiir Mas-chinenbau. Braunschweig Technische Universitat Carolo-Wilhelmina zu Braunschweig 2005. p. 119. 

The enzyme y3-galactosidase was purifled directly from crude cell extracts using expanded bed chromatography on another second generation matrix, Ni + loaded streamline chelating resin. The natural histidine content of the enzyme permitted the enzjune to be recovered in almost 90% jdeld and 5.95-fold purification from a crude unclarified cell extract (67). 

Non-soluble matrices with a controlled size of particles for expanded bed chromatography [11]. 

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