Chimeric polypeptides, polynucleotides encoding same, cells expressing same and methods of producing same
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 method of treating an inflammatory bowel disease in a subject in need thereof, the method comprising orally administering to the subject a therapeutically effective amount of plant cells comprising a nucleic acid construct having a nucleic acid sequence encoding 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,
wherein said therapeutically effective amount of said polypeptide in the plant cells comprises 0.02-0.27 mg/kg, thereby treating the TNFα-associated medical condition in the subject.
2. The method of claim 1 , wherein said therapeutically effective amount comprises 0.02-0.12 mg polypeptide/kg.
3. The method of claim 1 , wherein said therapeutically effective amount comprises 0.12-0.27 mg polypeptide/kg.
4. The method of claim 1 , wherein said inflammatory bowel disease is ulcerative colitis.
5. The method of claim 1 , wherein said inflammatory bowel disease is Crohn's disease.
6. The method of claim 1 , wherein said chimeric polypeptide comprises an additional domain encoding an endoplasmic reticulum signal peptide translationally fused N-terminally to said first domain.
7. The method of claim 6 , wherein said signal peptide is a plant signal peptide.
8. The method of claim 7 , wherein said plant signal peptide is as set forth in SEQ ID NO: 4.
9. The method of claim 1 , wherein said first domain is 200-250 amino acids long.
10. The method of claim 9 , wherein said first domain comprises the amino acid sequence LCAP (SEQ ID NO: 11) and VFCT (SEQ ID NO: 12).
11. The method of claim 10 , wherein said first domain further comprises the amino acid sequence LPAQVAFXPYAPEPGSTC (SEQ ID NO: 13).
12. The method of claim 11 , wherein said first domain is as set forth in SEQ ID NO: 2.
13. The method of claim 1 , wherein said immunoglobulin is IgG 1 .
14. The method of claim 1 , wherein said second domain is as set forth in SEQ ID NO: 9.
15. The method of claim 1 , wherein said chimeric polypeptide is as set forth in SEQ ID NO: 6.
16. The method of claim 1 , wherein said chimeric polypeptide comprises a plant-specific glycan.
17. The method of claim 1 , wherein said chimeric polypeptide comprises an additional domain encoding an endoplasmic reticulum retention signal peptide translationally fused C-terminally to said second domain.
18. The method of claim 1 , wherein said plant cells are tobacco cells from a suspension culture.
19. The method of claim 18 , wherein said tobacco plant cells from suspension culture are tobacco Bright Yellow (BY) 2 cells.
20. A method of treating an inflammatory bowel disease in a subject in need thereof, the method comprising orally administering to the subject a therapeutically effective amount of tobacco BY-2 cells from suspension culture recombinantly expressing a chimeric polypeptide comprising:
(i) a first domain which comprises SEQ ID NO: 2, and
(ii) a second domain which comprises SEQ ID NO: 9,
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,
wherein said therapeutically effective amount of said polypeptide in the plant cells comprises 0.02-0.27 mg/kg, thereby treating the TNFα-associated medical condition in the subject.
21. The method of claim 20 , wherein said chimeric polypeptide is as set forth in SEQ ID NO: 6.
22. The method of claim 20 , wherein said inflammatory bowel disease is ulcerative colitis.
23. The method of claim 20 , wherein said inflammatory bowel disease is Crohn's disease.
24. The method of claim 1 , wherein said chimeric polypeptide is as set forth in SEQ ID NO: 7.