METHOD FOR REPRESENTING THE PROTEZYM OF THE PROTEASE ACTIVATING THE BLOOD GENERATING FACTOR VII BY MEANS OF ION EXCHANGE CHROMATOGRAPHY

摘要

Purification of factor VII activating protease (I) or its proenzyme (II) from (biological) fluids produced by recombinant DNA methods comprising performing anion- and/or cation-exchange chromatography at a pH below the isoelectric point of (I) or (II), is new. Purification of factor VII activating protease (I) or its proenzyme (II) from (biological) fluids produced by recombinant DNA methods comprises anion- and/or cation-exchange chromatography at a pH below the isoelectric point of (I) or (II), optionally in combination with affinity chromatography and/or fractional precipitation at pH 2.5-9.0, where affinity chromatography is performed using calcium phosphate/hydroxyapatite, a hydrophobic matrix, a chelate matrix or a matrix coated with an immobilized mono- or polyclonal antibody or its Fab or F(ab)2 fragment, is new. An Independent claim is also included for a reagent for use in biological tests systems and antigen assays, comprising (I) and/or (II) and one or more protein stabilizers (III) such as: solubilizing agents, preferably hydroxyproline; detergents, preferably Tween (RTM) or Triton (RTM); proteins, e.g. albumin, gelatin, fibronectin or vitronectin; reducing agents, preferably dithiothreitol, mercaptoethanol or cysteine; protease inhibitors, e.g. aprotinin, alpha -2-antiplasmin, C1 esterase inhibitor, inter- alpha -trypsin inhibitor or antithrombin III/heparin; divalent ion complexing agents, preferably EGTA (undefined), ethylenediamine tetraacetatic acid (EDTA); divalent ions, preferably calcium ions; amino acids, preferably glutamate, arginine, lysine or glycine; sugars, preferably glucose, arabinose, mannose or mannitol; and/or alcohols, preferably ethylene glycol or polyethylene glycol