Rapid Purification Of Kallikrein With The Combination Of Hydrophobic Interaction Chromatography And Salt Precipitation

There are a number of purification methods of pancreas kallikrein, such as (NH4)2SO4 precipitation, ion exchange chromatography, organic solvent grading and affiny chromatography etc. At present, the purification based on pancreas kallikrein mostly adopts anion exchange chromatography in combination with cation exchange chromatography , adding a step of gel filtration as a final step. The pancreas kallikrein is eluted by changing the pH in the ion exchange process. Because of the buffer action, the pH changes slowly in the column and the process time is long, with consumption of a lot of salts. An example is Graham S. Bai ley' s purification. Though the purity of the pancreas kallikrein was up to 1254 u/mg through three purification steps, the recovery was only 28.4%.Combination of an ion exchange chromatography with a hydrophobic interaction chromatography or with an affinity chromatography has been practised in the literature. There are more steps in these processes, the purification cycle is long and the cost is relatively high, An example is the Talal S.EI.Thaher's purification course of English. Though the recovery was 87%, the purity of the pancreas kallikrein was just 156.3 u/mg through three purification steps.In this thesis, we combine hydrophobic interaction chromatography and (NH4)2SO4 precipitation to establish a novel process for purifying pancreas kallikrein. The method helps to shorten operating time, use less elution solution, maintain high protein activity and receive the higher recovery .The hydrophobic interaction chromatography needs the condition of high salt density to operate, and the salt is relatively high in the supernatant fluid after salting out in this...