C1 inhibitor produced in the milk of transgenic mammals

Abstract

The invention provides transgenic nonhuman mammals expressing C1 inhibitor in their milk. The C1 inhibitor is useful in treating patients with hereditary angioedema or patients requiring immunosuppression.

1 - 17 . (canceled)  
     
     
         18 . C1 inhibitor produced by 
 recovering milk from a transgenic nonhuman mammal comprising: a DNA segment encoding a C1 inhibitor heterologous to the mammal operably linked to at least one regulatory sequence effective to promote expression of the DNA segment in mammary gland cells of the mammal and a segment encoding a signal peptide functional in mammary cells of the mammal: wherein the DNA segment encoding the C1 inhibitor can be expressed in the mammary gland cells of an adult form of the mammal to produce C1 inhibitor in the milk of the nonhuman mammal; and    purifying the C1 inhibitor from the milk.    
     
     
         19 . The C1 inhibitor of  claim 18  that is a human C1 inhibitor, and which is free of other human proteins.  
     
     
         20 . A pharmaceutical composition comprising the C1 inhibitor of  claim 19  and a pharmaceutical carrier.  
     
     
         21 . The pharmaceutical composition of  claim 19 , wherein the pharmaceutical carrier is for intravenous administration.  
     
     
         22 . The pharmaceutical composition of  claim 21  that is sterile, substantially isotonic and produced under GMP conditions.  
     
     
         23 . A method of treating a patient suffering from or susceptible to C1 inhibitor deficiency, comprising administering to the patient an effective dose of C1 inhibitor produced in accordance with claim  1  and a pharmaceutically acceptable composition.  
     
     
         24 . A method of purifying human C1 inhibitor, comprising: loading a sample comprising human C1 inhibitor in milk from a nonhuman animal onto a cationic exchange column under conditions in which the human C1 inhibitor binds to the column; eluting the human C1 inhibitor from the cationic exchange column; loading the eluate on an anionic exchange column under conditions in which the human C1 inhibitor binds to the column; eluting the human C1 inhibitor from the anionic exchange column; loading the eluate onto a metal ion exchange column under conditions in which residual contaminating proteins bind to the column; collecting eluate containing the human C1 inhibitor from the metal ion exchange column.  
     
     
         25 . The method of  claim 24 , wherein the sample further comprises rabbit C1 inhibitor; and the eluate from the metal ion exchange column has a higher ratio of human C1 inhibitor to rabbit C1 inhibitor than the sample  
     
     
         26 . The method of  claim 25 , wherein the ratio is at least 500:1.  
     
     
         27 . A pharmaceutical composition for comprising human C1 inhibitor free of other human proteins.  
     
     
         28 . A pharmaceutical composition comprising human C1 inhibitor that is at least 98% w/w pure.  
     
     
         29 . The pharmaceutical composition of composition of  claim 27  that is sterile.  
     
     
         30 . A method of treating a patient suffering from or susceptible to C1 inhibitor deficiency, comprising administering to the patient an effective dose of a composition of  claim 27 .  
     
     
         31 - 32 . (canceled)