Methods of preparing a NELL peptide are disclosed.
대표청구항▼
What is claimed is: 1. A method of expressing a peptide in a mammalian cell, said method comprising: providing a nucleic acid construct including at least a nucleic acid encoding at least a nel-like (NELL) peptide in frame with a nucleic acid encoding a heterologous eukaryotic secretory signal pept
What is claimed is: 1. A method of expressing a peptide in a mammalian cell, said method comprising: providing a nucleic acid construct including at least a nucleic acid encoding at least a nel-like (NELL) peptide in frame with a nucleic acid encoding a heterologous eukaryotic secretory signal peptide; transfecting a mammalian cell with said nucleic acid construct; and culturing said mammalian cell under conditions that permit expression of the NELL peptide. 2. The method of claim 1, wherein the mammalian cell is a Chinese hamster ovary (CHO) cell. 3. The method of claim 2, wherein the nucleic acid encodes NELL1 or NELL2, wherein NELL1 is selected from the group comprising: SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6, wherein NELL2 is selected from the group consisting of: SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, and SEQ ID NO:14. 4. The method of claim 2, further comprising collecting NELL peptide secreted from the cell line; and substantially purifying the NELL peptide. 5. The method of claim 1, further comprising collecting the NELL peptide secreted from the cell line; and substantially purifying the NELL peptide. 6. The method of claim 4, further comprising testing the activity of the NELL peptide to induce bone formation. 7. The method of claim 5, further comprising testing the activity of the NELL peptide to induce bone formation. 8. The method of claim 4, wherein the purifying includes chromatography purification. 9. The method of claim 5, wherein the purifying includes chromatography purification. 10. A nucleic acid construct for expressing a nel-like (NELL) peptide in a mammalian cell, said nucleic acid construct comprising at least a nucleic acid encoding at least a NELL peptide in frame with a nucleic acid encoding a heterologous eukaryotic secretory signal peptide. 11. The nucleic acid construct of claim 10, wherein the nucleic acid encodes NELL1 or NELL2, wherein the NELL1 is selected from the group comprising: SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6, wherein NELL2 is selected from the group consisting of: SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, and SEQ ID NO:14. 12. The nucleic acid construct of claim 10, wherein the mammalian cell is a Chinese hamster ovary (CHO) cell. 13. A mammalian cell line for expressing a nel-like (NELL) peptide, said cell line including a nucleic acid construct comprising at least a nucleic acid encoding at least a NELL peptide in frame with a nucleic acid encoding a heterologous eukaryotic secretory signal peptide. 14. The cell line of claim 13, wherein the mammalian cell line comprises a Chinese hamster ovary (CHO) cell. 15. The cell line of claim 14, wherein the CHO cell secretes NELL1 or NELL2 peptide. 16. The cell line of claim 14, wherein the nucleic acid encodes NELL or NELL2 peptide, wherein NELL1 is selected from the group comprising: SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, or SEQ ID NO:6, and wherein NELL2 is selected from the group consisting of: SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, and SEQ ID NO:14.
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