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Protein purification with the KNAUER SMB equipment The KNAUER simulated moving bed (SMB) valve can operate with up to 16 columns assembled in parallel. Two different modes of operation are presented here. A) SMB chromatography SMB separation of binary protein mixtures Continuous process, continuous feed introduction into an 8, 12 or 16 columns arrangement, continuous generation of 2 value products (extract, raffinate) Columns rotate continuously, cyclic process, eluent consumption considerably reduced due to recycling. Examples: 1. 2. Separation of Myoglobin / Lysozyme with an aqueous gel filtration column environment Insulin purification B) High throughput/parallel chromatography Ion exchange chromatography (mostly anion exchange for proteins) All procedures can be performed in parallel which leads to a highly productive purification. The salt concentration from buffer 1-4 is stepwise increased. The proteins are selectively desorbed if the appropriate salt concentration passes through the column (step gradient). Columns 1-9 are loaded with proteins and the matrix is washed off (feed). Columns 10 and 11 are treated with buffer 1 to wash unwanted species away. Columns 12 and 13 are treated with buffer 2. Protein 1 is desorbed and collected. Columns 14 and 15 are free from protein 1 and treated with buffer 3. Protein 2 is desorbed and collected. Column 16 is reconditioned with buffer 4 to be ready for the next feeding. All columns move one position further after defined time intervals and return after one complete cycle back to their original positions. The systems can be additionally equipped with detectors (UV, conductivity, pH meter) according to the customer’s requirements. Examples: Purification of conalbumin, ovalbumin, myoglobin, transferrin, bovine serum albumin BUFFER 4 BUFFER 3 BUFFER 2 BUFFER 1 FEED 16 15 14 13 12 11 10 9 8 7 6 5 4 3 2 1 PROTEIN 2 WASH / WASTE PROTEIN 1 Wissenschaftliche Gerätebau Dr. Ing. Herbert Knauer GmbH Hegauer Weg 38, 14163 Berlin, Germany Phone: +49 30 809727-0 Fax: +49 30 8015010 E-Mail: [email protected] Internet. www.knauer.net PRODUCT / WASH / WASTE 2/2