Chromatographic separation processes are widely used to isolate and purify value added products. Most frequently, such separation processes are performed exploiting the principles of injecting samples of the feed mixture in a repetitive periodic manner and collecting the target products batchwise. In the early 1960s, an alternative operation principle based on using several columns connected in series and exploiting a continuous countercurrent movement between the mobile and stationary phases initiated significant improvements in performing specific separation processes in the petrochemical and sugar industries. In the last decade, the so-called simulated moving bed (SMB) chromatography has rapidly entered the pharmaceutical industry. In particular enantioseparations using chiral stationary phases have become a strong driving force to develop more sophisticated concepts of continuous chromatography. Currently, the growing interest in efficient methods for the downstream processing of biomolecules enhances interest in SMB processes. This paper gives a brief overview of the new suggestions and trends. © 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim [accessed July 4, 2008

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Background: Soybeans play a prominent role in allergologic research due to the high incidence of allergic reactions. For detailed studies on specific proteins it is necessary to have access to a large amount of pure substance.

Soya is considered to be one of the eight most significant food allergens. Among the allergenic soya proteins determined to date, P34 has been identified as one of the immunodominant soya antigens. Sensitisation to a specific food antigen like P34 generally follows the transit of intact antigens across the intestinal barrier and usually occurs in infants, who are most susceptible to food allergies. In the present study, we used the intestinal epithelial cell line IPEC-J2, which was originally derived from the jejunum of a neonatal piglet, to recapitulate the infant intestinal epithelium and study the binding and uptake of P34 protein. P34 was partially resistant to degradation in an in vitro proteolysis assay. IPEC-J2 cells were able to endocytose intact P34, as shown by immunofluorescence and immunoelectronmicroscopy methods. P34 associated with lipid raft microdomains of IPEC-J2 cells, and disruption of caveolae/lipid raft microdomains using methyl-b-cyclodextrin abolished P34 endocytosis, indicating that the observed endocytosis was mediated by caveolae. Using IPEC-J2 cells grown on Transwell filters, we further demonstrated that P34 is transported through the epithelial monolayer by transcytosis. Piglets frequently show hypersensitivity to soya antigens, and in this study, we show that healthy adult pigs with dietary exposure to soya protein mount an antibody response to soyabean protein P34, suggesting that this protein has entered the body, probably through gastrointestinal uptake. In summary, our data suggest that soya P34 resists proteolysis in the gastrointestinal tract and is transported through the intestinal epithelial barrier, thereby allowing sensitisation of immune cells in the sub-epithelial compartment.