Method for high-level secretory production of protein
AssigneesNational Institute of Advanced Industrial Science and Technology, Kyowa Hakko Kirin Co., Ltd., Daiichi Sankyo Company, Limited
InventorsYasunori Chiba, Yoshifumi Jigami, Yoshie Takahashi, Kosuke Kuroda, Kazuo Kobayashi, Kimihisa Ichikawa, Koichi Nonaka, Takeshi Suzuki, Minako Ono
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Abstract
This invention provides a means for high-level secretory production of a protein, and, in particular, a protein having a complicated structure such as an antibody, in a host cell such as a yeast cell. This invention provides a method for high-level secretory production of a foreign protein with the use of a transformed host cell having one or more types of chaperone protein genes and via suppression of O sugar chain inherent to a host cell such as a yeast cell.
Claims27
A transformed host cell which is transformed with a nucleic acid comprising an isolated polynucleotide encoding a protein disulfide isomerase (PDI), wherein said isolated polynucleotide is selected from the group consisting of (a) and (b):
- (a) an isolated polynucleotide comprising the nucleotide sequence of SEQ ID NO:
- 1;
- and
- (b) an isolated polynucleotide comprising a nucleotide sequence having at least 95% nucleotide sequence identity to the nucleotide sequence of SEQ ID NO:
- 1, wherein said nucleotide sequence having at least 95% nucleotide sequence identity to the nucleotide sequence of SEQ ID NO:
- 1 encodes a protein having an activity of accelerating increasing secretion of a foreign protein.
A transformed host cell which is transformed with a nucleic acid comprising an isolated polynucleotide encoding a protein disulfide isomerase (PDI), wherein said isolated polynucleotide encodes a polypeptide selected from the group consisting of (a) and (b):
- (a) a polypeptide comprising the amino acid sequence of SEQ ID NO:
- 2;
- and
- (b) a polypeptide comprising an amino acid sequence having at least 95% amino acid sequence identity to the amino acid sequence of SEQ ID NO:
- 2, wherein said polypeptide comprising an amino acid sequence having at least 95% amino acid sequence identity to the amino acid sequence of SEQ ID NO:
- 2 has an activity of increasing secretion of a foreign protein.
An isolated polynucleotide encoding a protein disulfide isomerase (PDI), wherein said isolated polynucleotide is selected from the group consisting of (a) and (b):
- (a) an isolated polynucleotide comprising the nucleotide sequence of SEQ ID NO:
- 1;
- and
- (b) an isolated polynucleotide comprising a nucleotide sequence having at least 95% nucleotide sequence identity to the nucleotide sequence of SEQ ID NO:
- 1, wherein said nucleotide sequence having at least 95% nucleotide sequence identity to the nucleotide sequence of SEQ ID NO:
- 1 encodes a protein having an activity of increasing secretion of a foreign protein in a recombinant host cell.
An isolated polynucleotide encoding a protein disulfide isomerase (PDI), wherein said isolated polynucleotide encodes a polypeptide selected from the group consisting of (a) and (b):
- (a) a polypeptide comprising the amino acid sequence of SEQ ID NO:
- 2;
- and
- (b) a polypeptide comprising an amino acid sequence having at least 95% amino acid sequence identity to the amino acid sequence of SEQ ID NO:
- 2, wherein said polypeptide comprising an amino acid sequence having at least 95% amino acid sequence identity to the amino acid sequence of SEQ ID NO:
- 2 has an activity of increasing secretion of a foreign protein in a recombinant host cell.
Dependencies are inferred from the claim text.