A supernatant from Phanerochaete chrysosporium cultivation was loaded on CIM® QA Disk, and elution was effected by a linear gradient at a flow rate of 3 mL/min (9 CV/min). Baseline separation of isoenzymes H2, H6/H7, H8 and H10 was achieved in less than 3 minutes.
50 mL of crude enzyme solution were loaded on CIM® DEAE 8 mL tube and eluted by a linear gradient at a flow rate of 1.2 mL/min. Analysis of the elution fraction resulted in 86.1% recovery and 4 times concentration of the sample, having a purity of >95%.
60 mL of non-concentrated periplasmic fraction from Prevotella bryantii cultivation medium were subjected to two-step purification on CIM® DEAE 8 mL tube. Both purification steps were performed under similar chromatographic conditions, i.e. same column, buffer systems and flow rate (7 mL/min). Gradient slopes varied to achieve isolation of the desired fraction. The elution fraction of interest of purification No1 was subjected to a second purification on the same column. By altering the linear gradient pure 66 kDa-Endoxylanase for production of antibodies was isolated.