Processes for immobilising biological entities

Abstract

According to the invention there is provided inter alia a process for the manufacture of a solid object having a surface comprising a layered coating of cationic and anionic polymer wherein the outer coating layer comprises an anticoagulant entity, comprising the steps of: i) treating a surface of the solid object with a cationic polymer; ii) treating the surface with an anionic polymer; iii) optionally repeating steps i) and ii) one or more times; iv) treating the surface with a cationic polymer; and v) treating the outermost layer of cationic polymer with an anticoagulant entity, thereby to covalently attach the anticoagulant entity to the outermost layer of cationic polymer; wherein, the anionic polymer is characterized by having (a) a total molecular weight of 650 kDa-10,000 kDa; and (b) a solution charge density of >4 μeq/g; and wherein, step ii) is carried out at a salt concentration of 0.25 M-5.0 M.

1 . A process for the manufacture of a solid object having a surface comprising a layered coating of cationic and anionic polymer wherein the outer coating layer comprises an anticoagulant entity, comprising the steps of:
 i) treating a surface of the solid object with a cationic polymer;   ii) treating the surface with an anionic polymer;   iii) optionally repeating steps i) and ii) one or more times;   iv) treating the surface with a cationic polymer; and   v) treating the outermost layer of cationic polymer with an anticoagulant entity, thereby to covalently attach the anticoagulant entity to the outermost layer of cationic polymer;   wherein,   the anionic polymer is characterized by having (a) a total molecular weight of 650 kDa-10,000 kDa; and (b) a solution charge density of >4 μeq/g;   and wherein,   step ii) is carried out at a salt concentration of 0.25 M-5.0 M.   
     
     
         2 . A process for the manufacture of a solid object according to  claim 1 , wherein the anionic polymer is dextran sulfate. 
     
     
         3 . A process for the manufacture of a solid object according to  claim 1 , wherein the anionic polymer is characterized by having a total molecular weight of 750 kDa-10,000 kDa. 
     
     
         4 . (canceled) 
     
     
         5 . A process for the manufacture of a solid object according to  claim 1 , wherein the anionic polymer is characterized by having a solution charge density of between >4 μeq/g and 7 μeq/g. 
     
     
         6 . A process for the manufacture of a solid object according to  claim 1 , wherein step ii) is carried out at a salt concentration of 0.25 M-4.0 M. 
     
     
         7 . (canceled) 
     
     
         8 . (canceled) 
     
     
         9 . A process for the manufacture of a solid object according to  claim 1 ,
 wherein the salt is selected from the group consisting of sodium chloride, sodium sulfate, sodium hydrogen phosphate and sodium phosphate.   
     
     
         10 . A process for the manufacture of a solid object according to  claim 9 , wherein the salt is sodium chloride. 
     
     
         11 . (canceled) 
     
     
         12 . (canceled) 
     
     
         13 . (canceled) 
     
     
         14 . A process for the manufacture of a solid object according to  claim 1 , wherein the cationic polymer of step i) is a polyamine. 
     
     
         15 . A process for the manufacture of a solid object according to  claim 1 , wherein the cationic polymer of step iv) is a polyamine. 
     
     
         16 . (canceled) 
     
     
         17 . (canceled) 
     
     
         18 . A process for the manufacture of a solid object according to  claim 1 , wherein the anticoagulant entity is a heparin moiety. 
     
     
         19 . A process for the manufacture of a solid object according to  claim 18 , wherein the heparin moiety is an end-point attached heparin moiety. 
     
     
         20 . A process for the manufacture of a solid object according to  claim 19 , wherein the end-point attached heparin moiety is connected through its reducing end. 
     
     
         21 . (canceled) 
     
     
         22 . (canceled) 
     
     
         23 . (canceled) 
     
     
         24 . (canceled) 
     
     
         25 . (canceled) 
     
     
         26 . (canceled) 
     
     
         27 . (canceled) 
     
     
         28 . (canceled) 
     
     
         29 . (canceled) 
     
     
         30 . (canceled) 
     
     
         31 . (canceled) 
     
     
         32 . (canceled) 
     
     
         33 . (canceled) 
     
     
         34 . (canceled) 
     
     
         35 . (canceled) 
     
     
         36 . (canceled) 
     
     
         37 . (canceled) 
     
     
         38 . (canceled) 
     
     
         39 . (canceled) 
     
     
         40 . (canceled) 
     
     
         41 . (canceled) 
     
     
         42 . (canceled) 
     
     
         43 . A solid object having a surface comprising a layered coating of cationic and anionic polymer, wherein the outer coating layer is a layer comprising cationic polymer to which is covalently bound an anticoagulant entity;
 and wherein the anionic polymer is characterized by having (a) a total molecular weight of 650 kDa-10,000 kDa; and (b) a solution charge density of >4 μeq/g.   
     
     
         44 . A solid object according to  claim 43 , wherein the anionic polymer is characterized by having a total molecular weight of 650 kDa-1,000 kDa. 
     
     
         45 . (canceled) 
     
     
         46 . (canceled) 
     
     
         47 . (canceled) 
     
     
         48 . A solid object according to  claim 43 , wherein the anionic polymer is applied to the surface at a salt concentration of 0.25 M-5.0 M. 
     
     
         49 . A solid object according to  claim 43 , wherein the anionic polymer is a polymer comprising groups selected from —CO 2   − , —SO 3   − , —PO 3 H −  and —PO 3   2− . 
     
     
         50 . A solid object according to  claim 49 , wherein the anionic polymer is a polymer comprising —SO 3   −  groups. 
     
     
         51 . A solid object according to  claim 50 , wherein the sulfur content of the anionic polymer is between 10% and 25% by weight of the anionic polymer. 
     
     
         52 . (canceled) 
     
     
         53 . (canceled) 
     
     
         54 . (canceled) 
     
     
         55 . (canceled) 
     
     
         56 . A solid object having a surface comprising a layered coating of cationic and anionic polymer, wherein the outer coating layer is a layer comprising cationic polymer;
 and wherein the anionic polymer is characterized by having (a) a total molecular weight of 650 kDa-10,000 kDa; and (b) a solution charge density of >4 μeq/g.   
     
     
         57 . (canceled) 
     
     
         58 . (canceled) 
     
     
         59 . (canceled) 
     
     
         60 . (canceled) 
     
     
         61 . A solid object having a surface comprising a layered coating of cationic and anionic polymer, wherein the outer coating layer is a layer comprising anionic polymer; and wherein the anionic polymer is characterized by having (a) a total molecular weight of 650 kDa-10,000 kDa; and (b) a solution charge density of >4 μeq/g. 
     
     
         62 . (canceled)