Affinity Chromatography of Liver Collagenase

概要

論文の詳細を見る
Affinity chromatography was done for identification of the liver collagenase.<BR>According to the Nagai's method, 0.5% acid soluble guinea pig skin collagen was entrapped into 5% acrylamide gel. The gel was ground to 10-50 mesh, and packed into 10×110mm column. The sample was applied through the column with pH 7.5 buffer at 4°, and eluted with pH 5.2 buffer. The recovery of the enzyme activity after affinity chromatography was usually extremely low and varied from 5 to 70%. The part of collagenase activity was passed through with pH 7.5, however, a sharp peak of collagenase activity without detectable protein absorption was recognized with pH 5.2 buffer, and the least amount of caseinolytic activity was determined in this fraction. Some collagenase samples from liver tissue culture medium, liver homogenate reacted with NaSCN, and separated Kupffer cells were purified by affinity chromatography and incubated with collagen. Those enzymes were recognized as collagenase from their mode of reaction by acrylamide disc electrophoresis.<BR>Affinity chromatography was seemed to be useful for detecting tissue collagenase, however, it was not seemed to be adequate to purify a large amount of the enzyme because of its small capacity, The reason for its inconstant recovery was attributed to the character of the gel entrapped collagen.
Japan Society of Clinical Chemistryの論文