Chimeric polypeptides, polynucleotides encoding same, cells expressing same and methods of producing same

Abstract

A plant produced chimeric polypeptide is provided. The plant produced chimeric polypeptide comprising: (i) a first domain which comprises a TNF Alpha binding domain of a TNF receptor, and (ii) a second domain which comprises an Fc domain of an immunoglobulin, wherein the first domain and the second domain are N-terminally to C-terminally respectively sequentially translationally fused and wherein the chimeric polypeptide specifically binds TNF Alpha.

1 . A plant produced chimeric polypeptide comprising:
 (i) a first domain which comprises a TNFα binding domain of a TNF receptor, and   (ii) a second domain which comprises an Fc domain of an immunoglobulin, wherein said first domain and said second domain are N-terminally to C-terminally respectively sequentially translationally fused and wherein the chimeric polypeptide specifically binds TNFα.   
     
     
         2 . A chimeric polypeptide comprising:
 (i) a first domain which comprises a TNFα binding domain of a TNF receptor;   (ii) a second domain which comprises an Fc domain of an immunoglobulin; and   (iii) a third domain comprising an endoplasmic reticulum retention signal, wherein said first domain, second domain and third domain are N-terminally to C-terminally respectively sequentially translationally fused and wherein the chimeric polypeptide specifically binds TNFα.   
     
     
         3 . The polypeptide of  claim 1  comprising an additional domain encoding an endoplasmic reticulum signal peptide translationally fused N-terminally to said first domain. 
     
     
         4 . The polypeptide of  claim 3 , wherein said signal peptide is a plant signal peptide. 
     
     
         5 . The polypeptide of  claim 4 , wherein said plant signal peptide is as set forth in SEQ ID NO: 4. 
     
     
         6 . The polypeptide of  claim 1 , wherein said first domain is 200-250 amino acids long. 
     
     
         7 . The polypeptide of  claim 6 , wherein said first domain comprises the amino acid sequence LCAP (SEQ ID NO: 11) and VFCT (SEQ ID NO: 12). 
     
     
         8 . The polypeptide of  claim 7 , wherein said first domain further comprises the amino acid sequence LPAQVAFXPYAPEPGSTC (SEQ ID NO: 13). 
     
     
         9 . The polypeptide of  claim 8 , wherein said first domain is as set forth in SEQ ID NO: 2. 
     
     
         10 . (canceled) 
     
     
         11 . The polypeptide of  claim 1 , wherein said second domain is as set forth in SEQ ID NO: 9. 
     
     
         12 . The polypeptide of  claim 2  as set forth in SEQ ID NO: 6. 
     
     
         13 . The polypeptide of  claim 2  as set forth in SEQ ID NO: 7, 204 or 205. 
     
     
         14 . The polypeptide of  claim 1 , wherein the polypeptide is purified to at least 98% homogeneity. 
     
     
         15 . The polypeptide of  claim 1 , capable of inhibiting TNFα-induced apoptosis. 
     
     
         16 . The polypeptide of  claim 1 , comprising a plant-specific glycan. 
     
     
         17 . (canceled) 
     
     
         18 . An isolated polynucleotide comprising a nucleic acid sequence encoding the polypeptide of  claim 1 . 
     
     
         19 - 20 . (canceled) 
     
     
         21 . A nucleic acid expression construct comprising a nucleic acid sequence encoding the polynucleotide of  claim 18 , and a cis-acting regulatory element active in a plant cell. 
     
     
         22 . (canceled) 
     
     
         23 . A plant cell comprising the nucleic acid construct of  claim 21 . 
     
     
         24 . The plant cell of  claim 23  being a  Nicotiana tabacum  plant cell. 
     
     
         25 . The plant cell of  claim 24 , wherein said  Nicotiana tabacum  L. cv plant cell is a Bright Yellow (BY-2) cell. 
     
     
         26 . The plant cell of  claim 23 , being lyophilized. 
     
     
         27 - 29 . (canceled) 
     
     
         30 . A method of treating a TNFα-associated medical condition in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the polypeptide of  claim 1 , thereby treating the TNFα-associated medical condition in the subject. 
     
     
         31 . A method of treating a TNFα-associated medical condition in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the plant cells of  claim 23 , thereby treating the TNFα-associated medical condition in the subject. 
     
     
         32 - 39 . (canceled) 
     
     
         40 . The method of  claim 31 , wherein said medical condition is an inflammatory bowel disease. 
     
     
         41 - 46 . (canceled) 
     
     
         47 . A method of producing a polypeptide comprising:
 providing a cell according to  claim 23 ; and   culturing said cell so as to produce said polypeptide.   
     
     
         48 - 50 . (canceled)