IP Library Granted Patent US 10,407,475
Granted Patent B2
US 10,407,475 · App. 15/307,322 · Granted Sep 10, 2019

Protein secretory factor with high secretory efficiency and an expression vector comprising the same

Inventors: Seung Hae Kim (Seoul, KR); Yeon Chul Kim (Seoul, KR); Saem Jung (Seoul, KR)
Assignee: LG CHEM, LTD.
C07K14/47C07K14/82C07K16/00C12N15/85C12P21/02C07K2317/51C07K2317/515C07K2319/00C07K2319/02
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Quick Facts
Patent No.
US 10,407,475
App. No.
15/307,322
Granted
Sep 10, 2019
Kind
B2
Abstract

The present invention relates to a novel protein secretion factor, a vector including a nucleic acid sequence encoding the protein secretion factor, and a transformed cell into which the vector is introduced. The invention also relates to a method of producing a target protein using the transformed cell including the vector.

Claims (22)

1. An expression cassette, comprising a promoter, which is operably linked to a nucleic acid sequence encoding a protein secretion factor having an amino acid sequence of SEQ ID NO: 2, which is operably linked to the gene encoding the target protein, wherein the protein secretion factor is not a natural signal sequence of the target protein.

2. A vector for secretory expression of target protein, comprising an expression cassette comprising a promoter, which is operably linked to a nucleic acid sequence encoding a protein secretion factor having an amino acid sequence of SEQ ID NO: 2, which is operably linked to a gene encoding a target protein, wherein the protein secretion factor is not a natural signal sequence of the target protein.

3. The vector according to claim 2 , wherein the protein is selected from the group consisting of human growth hormones, serum proteins, antibodies, immunoglobulins, cytokines, α-, β- and γ-interferons, colony-stimulating factors (GM-CSF), platelet-derived growth factors (PDGF), phospholipase-activating protein (PLAP)s, insulins, tumor necrosis factors (TNF), growth factors, hormones, calcitonins, calcitonin gene related peptides (CGRP), enkephalins, somatomedins, erythropoietins, hypothalamic secretion factors, prolactins, chorionic gonadotropins, tissue plasminogen activators, growth hormone releasing peptides (GHRP), thymic humoral factors (THF), asparaginases, arginases, arginine deaminases, adenosine deaminases, peroxide dismutases, endotoxinase, catalases, chymotrypsins, lipases, uricases, adenosine diphosphatases, tyrosinases, bilirubin oxidases, glucose oxidases, glucosidases, galactosidases, glucocerebrosidases, and glucourodinases.

4. A method of producing a target protein, comprising:

i) culturing a transformed cell, into which the vector according to claim 3 is introduced, to express a target protein and secrete the target protein to the outside of the cell; and

ii) recovering the target protein from a culture or culture supernatant of the cell of step i).

5. A transformed cell, comprising the vector according to claim 2 .

6. The transformed cell according to claim 5 , wherein the cell is an animal cell.

7. A method of producing a target protein, comprising:

i) culturing a transformed cell, into which the vector according to claim 2 is introduced, to express a target protein and secrete the target protein to the outside of the cell; and

ii) recovering the target protein from a culture or culture supernatant of the cell of step i).

8. The method according to claim 7 , further comprising purifying the recovered target protein.

9. The method according to claim 7 , wherein the cell is a Chinese Hamster Ovary (CHO) cell.

10. A vector for secretory expression of an antibody, wherein the vector comprises:

a) a first expression cassette comprising a promoter, which is operably linked to a nucleic acid sequence encoding a protein secretion factor having an amino acid sequence of SEQ ID NO: 1, which is operably linked to a gene encoding an antibody light chain; and

b) a second expression cassette comprising a promoter is operably linked to a nucleic acid sequence encoding a protein secretion having an amino acid sequence of SEQ ID NO: 3, which is operably linked to a gene encoding an antibody heavy chain.

11. The vector according to claim 10 , wherein the antibody light chain has an amino acid sequence of SEQ ID NO: 48, and the antibody heavy chain has an amino acid sequence of SEQ ID NO: 49.

12. A method of producing an antibody, comprising:

i) culturing a transformed cell, into which the vector according to claim 10 is introduced, to express an antibody and secrete the antibody to the outside of the cell; and

ii) recovering the antibody from a culture or culture supernatant of the cell of step i).

13. A transformed cell, comprising the vector according to claim 10 .

14. The transformed cell according to claim 13 , wherein the cell is an animal cell.

Assignments (2)
MERGER AND CHANGE OF NAME Recorded May 22, 2017
From: LG LIFE SCIENCES LTD.; LG CHEM, LTD.
To: LG CHEM, LTD.
Reel/Frame 042793/0501 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Dec 2, 2016
From: KIM, SEUNG HAE; KIM, YEON CHUL; JUNG, SAEM
To: LG LIFE SCIENCES LTD.
Reel/Frame 040498/0411 →
Priority Claims (1)
KR 10-2014-0052752 · Apr 30, 2014 · national
Continuity (1)
Related Publication 20170044224A1 · Feb 16, 2017
Cited By (1)
US 12,247,071