Optimized nucleic acid sequences coding for the alpha-chain of human chorionic gonadotropin

Patent metadata

Field	Value
Publication number	US-10464984-B2
Application number	US-201615566961-A
Country	US
Kind code	B2
Filing date	Apr 22, 2016
Priority date	Apr 24, 2015
Publication date	Nov 5, 2019
Grant date	Nov 5, 2019

Abstract

A host cell characterized in that it comprises integrated into its genome a sequence coding for the a chain of hCG, and use of the host cell to produce recombinant hCG.

First claim

The invention claimed is:

1. A polynucleotide sequence comprising an optimized hCG α-chain nucleic acid sequence selected from SEQ ID NO:1 and sequences having at least 97% sequence identity thereto, and SEQ ID NO:4 and sequences having at least 97% sequence identity thereto.

2. The polynucleotide sequence of claim 1 , comprising the nucleic acid sequence of SEQ ID NO:1.

3. The polynucleotide sequence of claim 1 , comprising a nucleic acid sequence having at least 97% sequence identity to SEQ ID NO:1.

4. The polynucleotide sequence of claim 1 , comprising the nucleic acid sequence of SEQ ID NO:4.

5. The polynucleotide sequence of claim 1 , comprising a nucleic acid sequence having at least 97% sequence identity to SEQ ID NO:4.

6. A cultured transformed cell having integrated into its genome an optimized hCG α-chain polynucleotide sequence comprising SEQ ID NO:1 or a sequence having at least 96.5% sequence identity thereto or SEQ ID NO:4 or a sequence having at least 96.5% sequence identity thereto.

7. The transformed cell according to claim 6 , wherein the integrated sequence comprises SEQ ID NO:1.

8. The transformed cell according to claim 6 , wherein the integrated sequence comprises a sequence having at least 96.5% sequence identity to SEQ ID NO:1.

9. The transformed cell according to claim 6 , wherein the integrated sequence comprises a sequence having at least 97% sequence identity to SEQ ID NO:1.

10. The transformed cell according to claim 6 , wherein the integrated sequence comprises SEQ ID NO:4.

11. The transformed cell according to claim 6 , wherein the integrated sequence comprises a sequence having at least 96.5% sequence identity to SEQ ID NO: 4 .

12. The transformed cell according to claim 6 , wherein the integrated sequence comprises a sequence having at least 97% sequence identity to SEQ ID NO: 4 .

13. The transformed cell according to claim 6 , wherein the transformed cell is the PER.C6 cell line deposited under ECACC No. 96022940 transformed to have integrated into its genome the optimized hCG α chain polynucleotide sequence.

14. The transformed cell according to claim 6 , further having integrated into its genome a cDNA encoding alpha-2,3-sialyltransferase.

15. The transformed cell according to claim 6 , further having integrated into its genome a nucleic acid sequence encoding the hCG β chain.

16. A method for producing recombinant protein in a cell, comprising culturing a cell according to claim 6 in a suitable medium, and harvesting the recombinant protein.

17. A method for producing recombinant hCG in a cell, comprising culturing a cell according to claim 13 in a suitable medium, and harvesting the recombinant protein.

Assignees

• Ferring Bv

Inventors

• Plaksin Daniel
• Grinhut Ayelet

Classifications

• C12N9/1081

  transferring other glycosyl groups (2.4.99) · CPC title

• C07K2317/41

  Glycosylation, sialylation, or fucosylation · CPC title

• C07K14/59 Primary

  Follicle-stimulating hormone [FSH]; Chorionic gonadotropins, e.g.hCG [human chorionic gonadotropin]; Luteinising hormone [LH]; Thyroid-stimulating hormone [TSH] · CPC title

• C07K2317/14

  Specific host cells or culture conditions, e.g. components, pH or temperature · CPC title

• A61P5/06

  of the anterior pituitary hormones, e.g. TSH, ACTH, FSH, LH, PRL, GH · CPC title