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Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification
Published on: September 21, 2011
Separation of the proteins present in pancreatic juice using hydrophobic interaction chromatography
1Department of Physiological Sciences, University of Manchester, United Kingdom.
Abstract:
We have previously described a method for separating pancreatic juice proteins by reversed-phase HPLC. However, the solvents used in such a system denature the proteins, whose characterization therefore depends solely on molecular weight determinations. We have therefore tested the suitability of hydrophobic interaction chromatography as an alternative method of separation. Using a TSK phenyl 5PW column with a decreasing, four-stage ammonium sulfate concentration gradient, it was possible to separate the major proteins in rat pancreatic juice. The identity of each protein was confirmed by measuring its molecular weight and by assaying its enzyme activity. Hydrophobic interaction chromatography represents an improved system for separating pancreatic secretory enzymes in active form.
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