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  • Purification of porcine pancreatic kallikrein using various affinity chromatographies and intestinal absorption of the purified kallikrein.

Purification of porcine pancreatic kallikrein using various affinity chromatographies and intestinal absorption of the purified kallikrein.

Advances in experimental medicine and biology (1983-01-01)
M Hirado, K Uchida, M Niinobe, S Fujii
ABSTRACT

For the simple purification of porcine pancreas kallikrein, various affinity chromatographies using L-prolyl-L-phenylalanyl-L-arginine (Pro-Phe-Arg-OH), tosyl-L-arginine (Tos-Arg-OH) and acetyl-L-phenylalanyl-L-arginine (Ac-Phe-Arg-OH) as ligands were examined. The purification was performed as follows using the extract from acetone powder of porcine pancreas as starting material; ammonium sulfate fractionation (30-80%), affinity chromatography (repeated) and gel filtration on Sephadex G-75, respectively. With these purification techniques, porcine pancreas kallikrein was purified about 2000-fold, respectively. From the results, these affinity chromatographies were considered to be effective for the purification of porcine pancreas kallikrein. Intestinal absorption of the purified kallikrein was also investigated using a rabbit. After the administration of the purified kallikrein into the intestinal tract, peripheral blood was taken from the ear vein and the plasma was assayed for Pro-Phe-Arg-7-amino-4-methylcoumarinamide hydrolytic activity. The concentration of the kallikrein in the blood was maximal at 3 hr after the administration and the absorption at this time amounted to about 0.07%.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Pro-Phe-Arg-7-amido-4-methylcoumarin, ≥95%