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本文结合胰脏多酶联产实验对胰蛋白酶抑制剂进行了探讨。制备了Ｔｒｙｐｓｉｎ－Ｓｅｐｈａｒｏｓｅ４Ｂ亲和层析柱，采用硫酸铵、三氯醋酸沉淀、离心、色谱、冷冻干燥等技术，对猪胰脏中胰蛋白酶抑制剂进行分离纯化，测定各步中的活性，对有活性的终产品进行ＳＤＳ－ＰＡＧＥ鉴定分析。亲和柱偶联率为９５．７６％，亲和率为９．５６ｍｇ蛋白／ｍｌ胶。从０．７Ｓ（ＮＨ４）２ＳＯ４沉淀溶解液中亲和吸附胰蛋白酶后液体中与从０．９Ｓ（ＮＨ４）２ＳＯ４上清中亲和吸附的胰蛋白酶抑制剂的色谱图同一般亲和色谱图，呈单一峰。而从０．９Ｓ（ＮＨ４）２ＳＯ４沉淀及三氯醋酸沉淀吸附胰蛋白酶后液体中吸附胰蛋白酶抑制剂的色谱图均有一小一大两个峰。吸附物解吸后经活性测定，仅０．９Ｓ沉淀上清液中有较高活性，而０．７Ｓ沉淀中活性低且不稳定，其余两种吸附物均无活性。各原液中未测到活性。用１ｋｇ胰脏制得１５ｍｇ胰蛋白酶抑制剂，酶比活为２０８９Ｔ．Ｉ．Ｕ／ｍｇ蛋白。该产品的电泳图谱显示，在电泳前沿和分子量为２０ＫＤ处同时有两条带。第一条带应为胰蛋白酶抑制剂。
In this paper, pancreatic multi-enzyme co-production experiments trypsin inhibitor. Trypsin-Sepharose4B affinity chromatography column was prepared. The trypsin inhibitor in porcine pancreas was isolated and purified by ammonium sulfate, trichloroacetic acid precipitation, centrifugation, chromatography and freeze-drying. The activity of each step was determined. The active final product was analyzed by SDS-PAGE. Affinity column affinity was 95.76% with affinity of 9.56 mg protein / ml gel. The chromatogram of the trypsin inhibitor affinity-adsorbed from 0.9S (NH4) 2SO4 supernatant from the 0.7S (NH4) 2SO4 precipitated solution shows the same affinity chromatogram as that of the trypsin- A single peak. However, the chromatograms of trypsin inhibitor adsorbed in trypsin after 0.9S (NH4) 2SO4 precipitation and trichloroacetic acid precipitation showed one small peak and two large peaks. After desorption of the adsorbate, the activity was measured in the supernatant of only 0.9S sedimentation, while the activities in the precipitation of 0.7S were low and unstable, and the other two adsorbates were inactive. No activity was detected in each stock solution. 15mg of trypsin inhibitor was made with 1kg of pancreas and the specific activity of the enzyme was 2089T. I. U / mg protein. The electropherogram of the product shows that there are two bands simultaneously at the electrophoresis front and at a molecular weight of 20KD. The first band should be trypsin inhibitor.