Cell adhesion resisting surfaces

Abstract

A coated surface that resists cell adhesion comprisesg hyaluronic acid directly bound to a plasma-treated polymer surface. A process for producing the coated surface is disclosed as are further modifications of the hyaluronic acid by attaching ligand-binding polypeptides (antibodies or antibody binding proteins).

What is claimed is:  
     
         1 . A process of coating a surface of an article with hyaluronic acid (HA) or alginic acid (AA) or a derivative of HA or AA, which consists essentially of the steps of 
 i) treating a surface of an article with a plasma that causes formation of nitrogen containing groups on said surface; and    ii) exposing the treated surface of the article to a solution containing hyaluronic acid or alginic acid, or a derivative thereof, in the presence of a carbodiimide and a reactive intermediate ester stabilizing compound;    to obtain a coated surface.    
     
     
         2 . The process of  claim 1  wherein the reactive intermediate ester stabilizing compound is selected from the group consisting of N-hydroxysuccinimide, hydroxysulfosuccinimide and hydroxybenzotriazolohydrate.  
     
     
         3 . The process of  claim 1  comprising the further step of allowing the coated surface to dry.  
     
     
         4 . The process of  claim 1  wherein the surface is selected from the group consisting of polystyrene, polyethylene, polypropylene, polyethylene terephthalate, polytetrafluoroethylene, polylactide and cellulose.  
     
     
         5 . The process of  claim 4  wherein the surface is polystyrene.  
     
     
         6 . The process of  claim 1  wherein step (ii) is carried out within 60 minutes of step (i).  
     
     
         7 . The process of  claim 1  wherein the plasma treatment is an NH 3  plasma treatment.  
     
     
         8 . The process of  claim 7  wherein the plasma treatment is carried out by placing the polymer article into the plasma chamber, evacuating the chamber to a 20 mTorr base pressure, establishing a 375 mTorr NH3 atmosphere, followed by a 25 sec plasma treatment.  
     
     
         9 . The process of  claim 1  wherein the concentration of HA is at least about 0.05% w/v.  
     
     
         10 . The process of  claim 9  wherein the concentration of HA is at least about 0.5% w/v.  
     
     
         11 . The process of  claim 1  wherein the ratio of carbodiimide and ester stabilizing compound-to-HA repeat unit is at least 1 and 0.5.  
     
     
         12 . A process of coating a plasma-treated surface of an article, which surface comprises nitrogen-containing groups, with HA or AA or a derivative of HA or AA, said process comprising: 
 exposing the plasma-treated surface to a solution of HA, AA or said derivative in the presence of a carbodiimide and a reactive intermediate stabilizing compound, to obtain a coated surface.    
     
     
         13 . A process of coating a plasma-treated surface of an article, which surface comprises nitrogen-containing groups, with HA or AA or a derivative of HA or AA, said process consisting essentially of: 
 exposing the plasma-treated surface to a solution of HA, AA or said derivative in the presence of a carbodiimide and a reactive intermediate stabilizing compound, to obtain a coated surface.    
     
     
         14 . The process of  claim 12  wherein the reactive intermediate ester stabilizing compound is selected from the group consisting of N-hydroxysuccinimide, hydroxysulfosuccinimide and hydroxybenzotriazolohydrate.  
     
     
         15 . The process of  claim 12  comprising the further step of allowing the coated surface to dry.  
     
     
         16 . The process of  claim 12  wherein the surface is selected from the group consisting of polystyrene, polyethylene, polypropylene, polyethylene terephthalate, polytetrafluoroethylene, polylactide and cellulose.  
     
     
         17 . The process of  claim 16  wherein the surface is polystyrene.  
     
     
         18 . The process of  claim 12  wherein the plasma treated surface is an NH3 plasma treated surface.  
     
     
         19 . The process of  claim 12  wherein the article is Primaria™ treated.  
     
     
         20 . The process of  claim 12  wherein the plasma treatment is carried out by placing the polymer article into the plasma chamber, evacuating the chamber to a 20 mTorr base pressure, establishing a 375 mTorr NH 3  atmosphere, followed by a 25 sec plasma treatment.  
     
     
         21 . The process of  claim 12  wherein the concentration of HA is at least about 0.05% w/v.  
     
     
         22 . The process of  claim 21  wherein the concentration of HA is at least about 0.5% w/v.  
     
     
         20 . The process of  claim 12  wherein the ratio of carbodiimide- and ester stabilizing compound-to-HA repeat unit is at least 1 and 0.5.  
     
     
         21 . A surface that is formed by the process of  claim 1 .  
     
     
         22 . A surface that is formed by the process of claims  12 .  
     
     
         23 . An article having the surface of  claim 21  or  22 .  
     
     
         24 . The article of  claim 23  that comprises a polymer.  
     
     
         25 . The article of  claim 24  that is selected from the group consisting of a petri dish, biological culture bottle or flask, a tissue slide, a multiwell plate, a foam, a fiber mesh, a scaffold.  
     
     
         26 . The article of  claim 25  that is a tissue slide having a plurality of wells.  
     
     
         27 . A surface formed by the process of  claim 1  that is resistant to organic solvents.  
     
     
         28 . A method for producing a cell-adhesion resistive (CAR) solid phase surface to which is covalently bonded at least a first ligand binding polypeptide, comprising the steps of: 
 (a) coating a polymer surface with HA, AA, or derivative in accordance with  claim 1;     (b) oxidizing said HA, AA or derivative to create an amine-reactive group; and    (c) exposing said oxidized HA, AA or derivative to a first ligand-binding polypeptide wherein covalent bonds are formed between amino groups of said polypeptide and said amine-reactive group, resulting in the covalent bonding of said polypeptide to the HA, AA or derivative,    thereby producing said CAR surface to which is covalently bonded first ligand-binding polypeptide.    
     
     
         29 . The method of  claim 28 , wherein 
 (i) said oxidizing step (b) is performed by providing an oxidizing agent that generates reactive aldehyde groups on said HA, AA or derivative, and    (ii) step (c) additionally comprises providing a reducing agent to said polypeptide and said surface that effects reductive amination that results in said covalent bond formation between said amino groups of said polypeptide and said reactive aldehyde groups.    
     
     
         30 . The method of  claim 28 , wherein, step (b) additionally comprises, either before step (c) or contemporaneously therewith, the step of converting carboxylate groups of said HA, AA or derivative to reactive esters by exposure to a carbodiimide and a reactive intermediate ester stabilizing compound  
     
     
         31 . The method of  claim 30  wherein the reactive intermediate ester stabilizing compound is selected from the group consisting of N-hydroxysuccinimide, hydroxysulfosuccinimide and hydroxybenzotriazolohydrate.  
     
     
         32 . The method of  claim 28 , further comprising, after step (c), 
 (d) contacting said covalently bonded first ligand binding polypeptide with a second ligand binding polypeptide that is a ligand for said first ligand binding polypeptide under conditions that result in the noncovalent binding of said second polypeptide to said first polypeptide.    
     
     
         33 . The method of  claim 29 , further comprising, after step (c), 
 (d) contacting said covalently bonded first ligand binding polypeptide with a second ligand binding polypeptide that is a ligand for said first ligand binding polypeptide under conditions that result in the noncovalent binding of said second polypeptide to said first polypeptide.    
     
     
         34 . The method of  claim 30 , further comprising, after step (c), 
 (d) contacting said covalently bonded first ligand binding polypeptide with a second ligand binding polypeptide that is a ligand for said first ligand binding polypeptide under conditions that result in the noncovalent binding of said second polypeptide to said first polypeptide.    
     
     
         35 . The method of  claim 28 , wherein said surface is selected from the group consisting of polystyrene, polyethylene, polypropylene, polyethylene terephthalate, polytetrafluoroethylene, polylactide and cellulose.  
     
     
         36 . The method of  claim 29 , wherein said oxidizing agent of step (b) is periodate.  
     
     
         38 . The method of  claim 28 , wherein said first ligand-binding polypeptide is (a) an antibody, (b) a receptor, (c) an immunoglobulin binding protein, (d) avidin or streptavidin, (e) a lectin, (f) a cell adhesion molecule or (f) an extracellular matrix protein or (g) a synthetic peptide.  
     
     
         39 . The method of  claim 38  wherein said first ligand-binding polypeptide is an immunoglobulin-binding protein selected from the group consisting of a native or recombinant staphylococcal protein A, a native or recombinant staphylococcal protein G, and recombinant protein A/G.  
     
     
         40 . The method of  claim 38  wherein said first ligand-binding polypeptide is (a) an antibody or antigen-binding fragment thereof, (b) an immunoglobulin binding protein, or (c) avidin or streptavidin.  
     
     
         41 . The method of  claim 32  wherein said second ligand-binding polypeptide is (a) an antibody or antigen-binding fragment thereof, (b) a receptor, (c) a lectin, (d) a cell adhesion molecule or (e) an extracellular matrix protein or (f) a synthetic peptide.  
     
     
         42 . The method of  claim 32  wherein 
 (a) said first ligand-binding polypeptide is protein A, protein G, or recombinant protein A/G; and  
 (b) said second ligand binding polypeptide is an antibody or antigen-binding fragment thereof.  
 
     
     
         43 . The method of  claim 32  wherein 
 (a) said first ligand-binding polypeptide is avidin or streptavidin; and  
 (b) said second ligand binding polypeptide is a biotinylated antibody.  
 
     
     
         44 . The method of any of  claim 28  wherein said first ligand-binding polypeptide is a anti-CD34 monoclonal antibody.  
     
     
         45 . The method of  claim 40  wherein said first ligand-binding polypeptide is a anti-CD34 monoclonal antibody.  
     
     
         46 . The method of  claim 38 , wherein said extracellular matrix polypeptide is selected from the group consisting of collagen, laminin, fibronectin and thrombospondin 1, vitronectin, elastin, tenascin, aggrecan, agrin, bone sialoprotein, cartilage matrix protein, fibrinogen, fibrin, fibulin, a mucin, entactin, osteopontin, plasminogen, restrictin, serglycin, SPARC/osteonectin, versican, von Willebrand Factor, a cadherin, a connexin, and a selectin.  
     
     
         47 . The method of  claim 41 , wherein said extracellular matrix polypeptide is selected from the group consisting of a collagen, laminin, fibronectin and thrombospondin 1, vitronectin, elastin, tenascin, aggrecan, agrin, bone sialoprotein, cartilage matrix protein, fibrinogen, fibrin, fibulin, a mucin, entactin, osteopontin, plasminogen, restrictin, serglycin, SPARC/osteonectin, versican, von Willebrand Factor, a cadherin, a connexin, and a selectin.  
     
     
         48 . An article useful for binding a target cell or target molecule, which target cell or molecule comprises a target ligand, said article comprising; 
 (a) a CAR surface coated with a layer comprising HA, AA, or a derivative of HA or AA;    (b) covalently bonded to said HA, AA or derivative, a first ligand-binding polypeptide that is capable of binding to said target ligand.    
     
     
         49 . The article of  claim 48  wherein said first ligand-binding polypeptide is (a) an antibody, (b) a receptor, (c) an immunoglobulin binding protein, (d) avidin or streptavidin, (e) a lectin, (f) a cell adhesion molecule, (f) an extracellular matrix peptide or polypeptide, or (g) a synthetic peptide.  
     
     
         50 . The article of  claim 49  wherein said first ligand-binding polypeptide is an immunoglobulin-binding protein selected from the group consisting of a native or recombinant staphylococcal protein A, a native or recombinant staphylococcal protein G, and recombinant protein A/G.  
     
     
         51  The article of  claim 48 , further comprising, bound to said covalently bonded first ligand binding polypeptide, 
 (c) a second ligand binding polypeptide that: 
 (i) comprises a target ligand for said first ligand binding polypeptide, and  
 (ii) is capable of binding to a target ligand of a target cell or target molecule.  
 
 
     
     
         52 . The article of  claim 51  wherein said second ligand-binding polypeptide is (a) an antibody or antigen-binding fragment thereof, (b) and immunoglobulin-binding protein (c) a receptor, (d) a lectin, (e) a cell adhesion molecule, (f) an extracellular matrix peptide or polypeptide or (f) a synthetic peptide.  
     
     
         53 . The article of  claim 52  wherein 
 (a) said first ligand-binding polypeptide is Staphylococcal protein A, Streptococcal protein G, or a recombinant protein A/G; and  
 (b) said second ligand binding polypeptide is an antibody or antigen-binding fragment thereof.  
 
     
     
         54 . The article of  claim 52  wherein 
 (a) said first ligand-binding polypeptide is avidin or streptavidin; and  
 (b) said second ligand binding polypeptide is a biotinylated antibody.  
 
     
     
         55 . The article of  claim 48  wherein said first ligand-binding polypeptide is a monoclonal antibody specific for CD34 or an antigen-binding fragment or derivative thereof.  
     
     
         56 . The article of  claim 51  wherein said second ligand-binding polypeptide is a anti-CD34 monoclonal antibody or an antigen binding fragment or derivative thereof.  
     
     
         57 . The article of  claim 49  wherein said extracellular matrix polypeptide is selected from the group consisting of a collagen, laminin, fibronectin and thrombospondin 1, vitronectin, elastin, tenascin, aggrecan, agrin, bone sialoprotein, cartilage matrix protein, fibrinogen, fibrin, fibulin, a mucin, entactin, osteopontin, plasminogen, restrictin, serglycin, SPARC/osteonectin, versican, von Willebrand Factor, a cadherin, a connexin, and a selectin.  
     
     
         58 . The method of  claim 48  wherein said surface is selected from the group consisting of polystyrene, polyethylene, polypropylene, polyethylene terephthalate, polytetrafluoroethylene, polylactide and cellulose.