The Purification Of Kallikrein From Animal's Pancreas

Kallikrein has diverse physiological function, and because of this it has been extensively used in clinic. It was used to cure cardiovascular disease, male infertility, microcirculation disorderis, etal, and was high regared by medicine industry. The kallikrein which was used in clinic was from porcine pancreas mainly, and only a few came from urine of man. There were many years since human began to purify kallikrein from porcine pancreas, but it deserved further study in order to find a purification method which has high yield, convenient operation and short operation cycle time.Study indicated that bovine pancreas kallikrein could lower blood pressure and modulate the blood circulation also. So, it has significant economy value and clinical application value.One of the materials of kallikrein was frash porcine pancreas, so the purification process was: porcine pancreas, acetone powder, ammonium sulfate powder, and chromatography. In the experiment, three kinds of method were used to purify the kallikrein. They were Gel Filtration Chromatography, Hydrophobic Interaction Chromatography and Anion Exchange Chromatography. Finally, according to purification purpose and application potential in industry, one media was selected——QAE-Sephadex media, which was economical and practical. In one step theactivity could achieve 342.5U/mg. It was 34.52 folds from crude pancreatic acetone extract.The other material of kallikrein was crude pancreatic ammonium sulphate extract, so the purification process was: dialyse and chromatography. In the experiment, four kinds of method were used to purify the kallikrein. They were Gel Filtration Chromatography, Hydrophobic Interaction Chromatography, Anion Exchange Chromatography and Cation Exchange Chromatography. As the same, a purification process was selected——combining CM-Sepharose CL-6B and Butyl-Sepharose4FF, according to purification purpose and application potential in industry. The kallikrein was purified 10.22 folds when this process was used. The final specific activity of 667.0U/mg was obtained after two steps purification, and the yield was 51.6%.