Immunoadsorption of anti-von Willebrand Factor cleaving protease antibodies

Abstract

This invention relates to adsorbents and methods for highly selective removal of anti-von Willebrand Factor-cleaving protease antibodies (“anti-vWF-cp-abs”) from human plasma using human von Willebrand Factor-cleaving protease (“hvWF-cp”) or fragments thereof as affinity ligands. The adsorbents can be used for treating disorders associated with the occurrence of anti-vWF-cp-abs in patients, such as thromboembolic diseases.

We claim:  
     
         1 . An adsorbent for selective removal of anti-von Willebrand Factor-cleaving protease antibodies from human plasma, comprising human von-Willebrand Factor-cleaving protease or fragments thereof, coupled to an inert matrix material, said human von-Willebrand Factor-cleaving protease or fragments thereof having at least one functionally active epitope directed to said anti-von Willebrand Factor-cleaving protease antibodies under physiological conditions.  
     
     
         2 . The adsorbent according to  claim 1 , wherein said human von-Willebrand Factor cleaving protease is a recombinant human von-Willebrand Factor cleaving protease or fragments thereof.  
     
     
         3 . The adsorbent according to  claim 1 , wherein said matrix material is selected from the group consisting of carbohydrates, silicates, glasses and organic polyreaction products.  
     
     
         4 . The adsorbent according to  claim 3 , wherein said carbohydrates comprise cross-linked modified agarose.  
     
     
         5 . The adsorbent according to  claim 1 , wherein said matrix material is present in the form of porous beads.  
     
     
         6 . The adsorbent according to  claim 1 , wherein said von-Willebrand Factor-cleaving protease or fragments thereof are coupled directly or indirectly via a spacer to said matrix material.  
     
     
         7 . The adsorbent according to  claim 1 , wherein said fragments are selected from the group consisting of SEQ ID NOs:1-5, or from the group consisting of peptides having a length in the range of from about 6 to about 30 aa.  
     
     
         8 . An adsorbent for selective removal of anti-von Willebrand Factor-cleaving protease antibodies from human plasma, comprising recombinant human von-Willebrand Factor-cleaving protease or fragments thereof coupled to beads consisting of cross-linked modified porous agarose as inert matrix material, said recombinant human von-Willebrand Factor-cleaving protease or fragments thereof having at least one functionally active epitope directed to said anti-von Willebrand Factor-cleaving protease antibodies under physiological conditions.  
     
     
         9 . A method for selective removal of anti-von Willebrand Factor-cleaving protease antibodies from human plasma, comprising the steps of: 
 (a) providing a plasma from whole blood of a patient having a disorder associated with occurrence of anti-von Willebrand Factor-cleaving protease antibodies;    (b) subjecting said plasma to an affinity chromatography using an adsorbent comprising human von-Willebrand Factor-cleaving protease or fragments thereof, coupled to an inert matrix material, said human von-Willebrand Factor-cleaving protease or fragments thereof having at least one functionally active epitope directed to said anti-von Willebrand Factor-cleaving protease anti-bodies under physiological conditions; and    (c) collecting the flow-through fraction which is essentially devoid of anti-von Willebrand Factor-cleaving protease antibodies.    
     
     
         10 . The method according to  claim 9 , wherein said human von-Willebrand Factor cleaving protease is a recombinant human von-Willebrand Factor cleaving protease or fragments thereof.  
     
     
         11 . The method according to  claim 9 , wherein in step (b) PBS is used as a binding buffer.  
     
     
         12 . The method according to  claim 9 , wherein, after step (c), the adsorbent is regenerated by eluting the bound antibodies and re-equilibrating in PBS.  
     
     
         13 . The method according to  claim 9 , wherein the disorder is a thromboembolic disease.  
     
     
         14 . The method according to  claim 13 , wherein the thromboembolic disease is selected from the group consisting of thrombotic thrombocytic purpura, Henoch-Schonlein purpura, preeclampsia, neonatal thrombocytopenia or hemolytic-uraemic syndrome.  
     
     
         15 . The method according to  claim 9 , wherein said matrix material is selected from the group consisting of carbohydrates, silicates, glasses and organic polyreaction products.  
     
     
         16 . The method according to  claim 9 , wherein said carbohydrates comprise cross-linked modified agarose.  
     
     
         17 . The method according to  claim 9 , wherein said matrix material is present in the form of porous beads.  
     
     
         18 . The method according to  claim 9 , wherein said fragments are selected from the group consisting of SEQ ID NOs:1-5, or from the group consisting of peptides having a length in the range of from 6 to about 30 aa.  
     
     
         19 . A method for selective removal of anti-von Willebrand Factor-cleaving protease antibodies from human plasma, comprising the steps of: 
 (a) providing a plasma from whole blood of a patient having a disorder associated with occurrence of anti-von Willebrand Factor-cleaving protease antibodies;    (b) subjecting said plasma to an affinity chromatography using an adsorbent comprising recombinant human von-Willebrand Factor-cleaving protease or fragments thereof coupled to beads consisting of cross-linked modified porous agarose as inert matrix material, said recombinant human von-Willebrand Factor-cleaving protease or fragments thereof having at least one functionally active epitope directed to said anti-von Willebrand Factor-cleaving protease anti-bodies under physiological conditions, in PBS as binding buffer; and    (c) collecting the flow-through fraction which is essentially devoid of anti-von Willebrand Factor-cleaving protease antibodies.    
     
     
         20 . A method for selective removal of anti-von Willebrand Factor-cleaving protease antibodies from human plasma, comprising the steps of: 
 (a) providing a plasma from whole blood of a patient having a disorder associated with occurrence of anti-von Willebrand Factor-cleaving protease antibodies;    (b) subjecting said plasma to an affinity chromatography using an adsorbent comprising recombinant human von-Willebrand Factor-cleaving protease of fragments thereof coupled to beads consisting of cross-linked modified porous agarose as inert matrix material, said recombinant human von-Willebrand Factor-cleaving protease or fragments thereof having at least one functionally active epitope directed to said anti-von Willebrand Factor-cleaving protease anti-bodies under physiological conditions, in PBS as binding buffer;    (c) collecting the flow-through fraction which is essentially devoid of anti-von Willebrand Factor-cleaving protease anti-bodies;    wherein, after step (c), the adsorbent is regenerated by eluting the bound antibodies and re-equilibrating in PBS.