Method Of Complexing A Protein By The Use Of A Dispersed System And Proteins Thereof

Abstract

The present invention relates to methods for complexing a protein in a dispersed medium. Also disclosed are associated proteins produced by the methods of complexing of the present invention. Pharmaceutically effective stabilized protein dosages are also disclosed. The present invention also relates to a method for associating AHF protein in a dispersed medium.

1 . A method for complexing a protein in a dispersed medium, said method comprising:
 providing a protein;   altering a conformational state of the protein to expose hydrophobic domains therein by contacting the protein with a mixture comprising ethanol at a concentration of about 10-50%;   binding a stabilizer to the exposed hydrophobic domains; and   at least partially reversing said alteration to associate at least a portion of the protein with the stabilizer.   
   
   
       2 . The method according to  claim 1 , wherein the mixture has a pH range from about 7.0 to about 7.4. 
   
   
       3 . The method according to  claim 1 , wherein said contacting is carried out at a temperature of about 40° C. to about 74° C. 
   
   
       4 . The method according to  claim 3 , wherein said contacting is carried out at a temperature of about 70° C. to about 74° C. 
   
   
       5 . The method according to  claim 3 , wherein said contacting is carried out at a temperature of about 58° C. to about 62° C. 
   
   
       6 . The method according to  claim 3 , wherein said contacting is carried out at a temperature of about 40° C. 
   
   
       7 . The method according to  claim 1 , wherein the protein is interferon-gamma. 
   
   
       8 . The method according to  claim 1 , wherein the protein is Factor VIII. 
   
   
       9 . The method according to  claim 8 , wherein the protein is AHF. 
   
   
       10 . The method according to  claim 1 , wherein said reversing comprises solvent removal. 
   
   
       11 . The method according to  claim 1 , wherein said reversing comprises dialysis. 
   
   
       12 . The method according to  claim 1 , wherein said altering comprises unfolding the protein. 
   
   
       13 . The method according to  claim 1 , wherein said altering comprises stabilizing secondary or substantially secondary protein structure(s) of the protein. 
   
   
       14 . The method according to  claim 1 , wherein said reversing comprises refolding the protein. 
   
   
       15 . The method according to  claim 1 , wherein the mixture is an ethanol-water mixture. 
   
   
       16 . The method according to  claim 1 , wherein the mixture is an ethanol-phosphate buffered saline solution mixture. 
   
   
       17 . The method according to  claim 1 , wherein the ethanol is denatured ethanol. 
   
   
       18 . The method according to  claim 17 , wherein the ethanol is 190% proof grade ethanol or 200% proof grade ethanol. 
   
   
       19 . A method for complexing AHF protein in a dispersed medium, comprising:
 providing an AHF protein;   altering the conformational state of said AHF protein to expose hydrophobic domains therein;   binding a stabilizer to said exposed hydrophobic domains; and   at least partially reversing said alteration to associate at least a portion of said protein with said stabilizer.   
   
   
       20 . A method for complexing a protein in a dispersed medium, said method comprising:
 providing a protein;   altering a conformational state of the protein to expose hydrophobic domains therein;   binding a stabilizer to said exposed hydrophobic domains; and   at least partially reversing said alteration to associate at least a portion of the protein with the stabilizer.