Expanded-bed adsorption

Another application shows the preparative purification and polishing of a therapeutic fusion protein with a humanized recombinant IgG protein. The fusion protein was expressed by the fermentation of baby hamster kidney cells. The filtered culture supernatant (155 liters) contained 2.2 g of IgG and 75.5 g of total protein. After the immunoglobulins were isolated by expanded bed adsorption and rebuffering, the IgG fraction was bound to Fractogel EMD SOj (M). This column achieved baseline separation of complete antibodies (fusion protein) from small amounts of antibodies lacking the fusion part. The resulting highly purified IgG fraction (110 ml) was diluted to 150 ml and... 

The degree of bed expansion contributes to the efficiency of fluidised bed/expanded bed adsorption as a composite function of liquid distribution, liquid and particle properties (size, shape and density) and process conditions. Besides being an important design feature, the degree of bed expansion may be used as a quick and simple measure of bed stability.48... 

Bermejo, R. et al.. Preparative purification of B-phycoerythrin from the microalga Porphyridium cruentum by expanded-bed adsorption chromatography, J. Chromatogr. B, 790, 317, 2003. 

Bermejo, R. et ah. Expanded bed adsorption chromatography for recovery of phyco-cyanins from the microalga Spirulina platensis, Chromatographia, 63, 59, 2006. Thieghem, (van) R, Monascus, genre nouveau de I ordre des ascomycfetes. Bull. Soc. Bot. France, 31, 226, 1884. 

Expanded-bed adsorption (ERA) has gained popularity in bioprocessing since its commercial introduction in the 1990s because of its ability to handle a crude feedstock that contains cells or other particulates. ERA ehminates the need for a dedicated clarification step by combining sohd-hquid separation and adsorption into a single-unit operation [Hjorth et al., in Subramanian (ed.), op. cit., vol. 1, pp. 199-226 Mattiasson et al., in Ahuja (ed.), op. cit., pp. 417-451]. A... 

Clemmit, R.H. and Chase, H.A., Immobilized metal affinity chromatography of P-galactosidase from unclarified Escherichia coli homogenates using expanded bed adsorption, /. Chromatogr. A, 874, 27, 2000. 

Feuser, J., Halfar, M., Liitkemeyer, D., Ameskamp, N., Kula, M.-R., and Thommes, J., Interaction of mammalian cell culture broth with adsorbents in expanded bed adsorption of monoclonal antibodies, Process Biochem., 34,159,... 

G. Wang, Isolation and purification of phycoeryhtrin from red alga Gracilaria verrucosa by expanded-bed-adsorption and ion-exchange chromatography. Chromatogaphia 56 (2002) 509-513. 

Inulinase Candida kefyr Comparison of cross-flow filtration, RME, expanded-bed adsorption techniques for DSP [88]... 

Lihme, A. O. F., Oehlenschlaeger, R., Olsen, B. A., Zafirakos, E. (2003). Expanded bed adsorption system. United States Patent 6620326 (http //www.patentstorm.us/patents/6620326-description.html). 

Lokra, S., Helland, M. H., Claussen, I. C., Straetkvern, K. O., Egelandsdal, B.10.1016/j.lwt.2007.07.006 (2007). Chemical characterization and functional properties of a potato protein concentrate prepared by large-scale expanded bed adsorption chromatography. LWT - Food Sci. Technol. 

Strffitkvern, K. O., Schwarz, J. G., Wiesenborn, D. R, Zafirakos, E., Lihme, A. (1999). Expanded bed adsorption for recovery of patatin from crude potato juice. Bioseparation, 7, 333-345. 

If the product is secreted, recovery may involve a simple filtration step to remove any cells and cellular debris. Other clarification techniques include centrifugation and expanded bed adsorption [25], For such intracellular products as recombinant proteins produced in E. coli, the product may be denatured and located in inclusion bodies within the cells [26], Bacterial cells are typically concentrated by centrifugation or crossflow filtration, washed, and then disrupted by homogenization. Inclusion bodies are then isolated, and the protein product extracted and refolded. Validation of recovery operations for an E. coli product is described by Seely et al. [27],... 

Lindgren A, Johannson S, Nystrom L-E (1993) Scale-up of expanded bed adsorption. In Henon B (ed) BED - The seventh Bioprocess Engineering Symposium. The American Society of Mechanical Engineers, p 27... 

A number of applications are given in this chapter that detail the capture and recovery of intracellular proteins including recombinant proteins. The initial protein recovery steps, regardless of the source, are usually associated with large volumes and crude solutions, requiring removal of particles and reduction of volume before their purification can take place. Centrifugation, filtration, precipitation, solvent extraction, and batch adsorption are common unit operations involved in the preliminary steps of protein recovery. Expanded-bed adsorption, as described here, is an approach for the initial protein recovery that eliminates the need for clarification and volume reduction. In this process, a crude starting solution is pumped directly onto an... 

PHYSICOCHEMICAL BASIS OF EXPANDED-BED ADSORPTION FOR PROTEIN PURIFICATION... 

Expanded-bed adsorption has been used in a number of processes mainly for the recovery of protein from feedstocks containing cells or cell debris. It has been reviewed recently.19 However, expanded-bed processing has also gained attention in other areas of biotechnology such as bioremediation. Immobilized expanded bed technology can also be applied in analytical applications. 

The expanded-bed adsorption procedure was successfully applied for the recovery of both intracellular and extracellular protein from crude feedstock containing particulate matter. The technology has been applied by many industries both at the laboratory scale and at the pilot plant level for the recovery of several different recombinant proteins. 

Monoclonal antibodies were purified using immobilized protein A in expanded-bed mode.30 It was possible to capture 2 g of pure mouse IgG 2a per cycle from an unclarified hybridoma cell culture, by using a 5 cm diameter expanded-bed column. Recently recombinant proteins have been recovered from milk of transgenic live stock in a one-step expanded-bed adsorption process.31 Other applications of expanded bed purification of proteins are reviewed recently.19 Table 232 35 lists some relevant applications... 

Bruce, L. J., and Chase, H. A. (1998). The effect of column alignment on separation efficiency in scaled down expanded bed adsorption. Int. Conf. Expanded Bed Adsorption, 2nd, Napa Valley, CA, 1998, Abstr. P7 3. 

Hjorth, R. (1997). Expanded bed adsorption in industrial bioprocessing Recent developments. Trends Biotechnol. 15, 230-235. 

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