Claims for Patent: 7,045,318
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Summary for Patent: 7,045,318
Title: | Recombinant fusion proteins to growth hormone and serum albumin |
Abstract: | The invention describes methods of producing fusion proteins of albumin and growth hormone and products obtained by the methods. |
Inventor(s): | Ballance; David James (Nottingham, GB) |
Assignee: | Delta Biotechnology Limited (GB) |
Application Number: | 09/984,010 |
Patent Claims: | 1. A method of producing a fusion protein of serum albumin and growth hormone comprising: (a) expressing from a host cell said fusion protein, wherein said host cell
comprises a PRB1 promoter element operatively associated with a polynucleotide encoding said fusion protein; and (b) recovering said protein.
2. The method of either claim 1, wherein said polynucleotide is further operatively associated with a termination sequence. 3. The method of claim 2, wherein said termination sequence is S. cerevisiae ADH1 termination sequence. 4. A method of producing a fusion protein of serum albumin and growth hormone comprising: (a) expressing from a host cell said fusion protein, wherein said host cell comprises a polynucleotide encoding said fusion protein operatively associated with a S. cerevisiae ADH1 termination sequence; and (b) recovering said protein. 5. The method of claim 4, wherein said host cell further comprises a promoter of a gene operatively associated with said polynucleotide, wherein said promoter is selected from: (a) GAL1; (b) GAL10; (c) CYC1; (d) PHO5; (e) TRP1; (f) ADH1; (g) ADH2; (h) glyceraldehyde-3-phosphate dehydrogenase; (i) hexokinase; (j) pyruvate decarboxylase; (k) phosphofructokinase; (l) triose phosphate isomerase; (m) phosphoglucose isomerase; (n) glucokinase; (o) .alpha.-mating factor pheromone; (p) GUT2; (q) GPD1; (r) nmt; (s) fbp1; (t) AOX1; (u) AOX2; (v) MOX1; (w) FMD1; or (x) PGK1. 6. The method of claim 4, wherein said host cell further comprises a PRB1 promoter element operatively associated with said polynucleotide. 7. The method of either claim 1 or 4, wherein said polynucleotide further encodes a leader sequence, and said fusion protein comprises mature human serum albumin and mature human growth hormone. 8. The method of claim 7, wherein said leader sequence is an HSA/MF.alpha.-1 hybrid leader sequence. 9. The method of claim 7, wherein said leader sequence is selected from: (a) an invertase leader sequence; (b) an acid phosphatase leader sequence; (c) a pre-sequence of MF.alpha.-1; (d) a pre-sequence of .beta.-glucanase; (e) a pre-sequence of killer toxin; (f) a glucoamylase II leader sequence; (g) an .alpha.-galactosidase leader sequence; (h) a killer toxin leader sequence; or (i) a glucoamylase leader sequence. 10. A method of producing a fusion protein of serum albumin and growth hormone comprising: (a) expressing from a host cell said fusion protein, wherein said host cell comprises a PRB1 promoter element and S. cerevisiae ADH1 termination sequence operatively associated with a polynucleotide encoding: (1) an HSA/MF.alpha.-1 hybrid leader sequence and (2) said fusion protein; and (b) recovering said protein. 11. The method of any one of claims 1, 4, or 10, wherein said serum albumin is human serum albumin. 12. The method of claim 11, wherein said human serum albumin is the mature human serum albumin. 13. The method of any one of claims 1, 4, or 10, wherein said growth hormone is human growth hormone. 14. The method of claim 13, wherein said human growth hormone is the mature human growth hormone. 15. The method of claim 14, wherein said serum albumin is mature human serum albumin. 16. The method of any one of claims 1, 4, or 10, wherein said host cell is a yeast. 17. The method of claim 16, wherein said yeast is S. cerevisiae. 18. The method of claim 16, wherein said yeast is selected from: (a) S. italicus; (b) S. rouxii; (c) K. fragills; (d) K. lactis; (e) K. marxianus; (f) T. delbrueckil; (g) P. angusta; (h) P. anomala; or (i) P. pastoris. 19. The method of any one of claims 1, 4 or 10, wherein the host cell is selected from: (a) a bacterial cell; (b) an animal cell; (c) a fungal cell; (d) a plant cell; or (e) an insect cell. 20. The method of any one of claims 1, 4, or 10, wherein said host cell comprises a vector containing said polynucleotide encoding said fusion protein. 21. The method of any one of claims 1, 4, or 10, wherein the C-terminus of said serum albumin is fused to the N-terminus of said growth hormone. 22. The method of any one of claims 1, 4, or 10, wherein the N-terminus of said serum albumin is fused to the C-terminus of said growth hormone. 23. The method of any one of claims 1, 4, or 10, wherein said polynucleotide further encodes a flexible linker sequence introduced between said serum albumin and said growth hormone of said fusion protein. 24. The method of claim 23, wherein said flexible linker sequence comprises repeating units of [Gly-Gly-Gly-Gly-Ser].sub.2 (SEQ ID NO:16) or [Gly-Gly-Gly-Gly-Ser].sub.3 (SEQ ID NO:17). 25. The product obtained by the method of any one of claims 1, 4, or 10. |
Details for Patent 7,045,318
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