Protease activity determination in biological samples and liquid test sample, e.g. blood sample and blood serum, involves withdrawing protease from test sample of protease inhibitor by bringing test sample in contact with support material

摘要

Protease activity determination involves withdrawing the protease from the test sample of the protease inhibitor by bringing the test sample in contact with a support material. An inhibitor-binding substance linked to the support material covalently or by adsorption, has high affinity or bond strength for the protease inhibitor than the protease. The support material bounded with the protease inhibitor is separated and a substrate for the protease, whose activity is to be determined, is added. The proteolytic reaction of the substrate takes place by the protease. Independent claims are included for: (1) a device for determining the activity of enzymes, particularly for executing the procedure for determining activity of protease in a liquid test sample, which comprises an equipment, e.g. a chromatography column, for withdrawing the inhibitor, where the equipment contains a support material and a substance bound to the support material; and (2) a kit for executing the procedure for determining activity of enzymes and protease in a liquid test sample, which has 7-amino-4-trifluoromethyl coumarin (AFC) substrate in pure form or dissolved in a measuring buffer, solid AFC as calibration substance, support material with papain, which is bounded covalently or by adsorption, and specific, synthetic inhibitor, e.g. E64 inhibitor for cysteine proteases or CA074 inhibitor cahepsin B.