Method for production of polypeptide
AssigneeChugai Seiyaku Kabushiki Kaisha
InventorsHisahiro Tabuchi, Satoshi Tainaka, Tomoya Sugiyama
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Abstract
The present invention provides a method capable of producing a natural or recombinant protein in high yield. The present invention relates to a method of producing a polypeptide, comprising culturing a cell which strongly expresses cysteine sulfinic acid decarboxylase and has a transferred DNA encoding a desired polypeptide and thereby allowing the cell to produce the polypeptide. Hamster cysteine sulfinic acid decarboxylase, a DNA encoding the same, a recombinant vector and a transformed cell are also provided.
Claims13
A method for producing a desired antibody, in which cells capable of high-yield production of β-alanine and into which DNA encoding the desired antibody has been transferred arc cultured to produce the desired antibody, wherein the cells capable of high-yield production of β-alanine are cells into which DNA encoding cysteine sulfinic acid decarboxylase or a mutant thereof has been transferred, wherein the mutant has the same enzyme activity as cysteine sulfinic acid decarboxylase.
The production method of claim I, wherein the cells capable of high-yield production of β-alanine are capable of increasing the β-alanine concentration in the culture medium on 6th or subsequent days after the initiation of the culture by 50 μM or more.
A method tbr producing a desired antibody, in which cells that strongly express cysteine sulfinic acid decarboxylase and into which DNA encoding the desired antibody has been transferred are cultured to produce the desired antibody, wherein the cells that strongly express cysteine sulfinic acid decarboxylase are cells into which DNA encoding. cysteine sulfinic acid decarboxyiase has been transferred.
A recombinant vector comprising an isolated DNA encoding cysteine sulfinic acid decarboxylase, which is any one of the following (a) to (e), provided that DNA having the nucleotide sequence as shown in SEQ ID NO:
- 3, DNA having the nucleotide sequence as shown in SEQ ID NO:
- 5, and DNA having the nucleotide sequence as shown in SEQ ID NO:
- 7 are excluded:
- (a) a DNA encoding a polypeptide having the amino acid sequence as shown in SEQ ID NO:
- 2;
- (b) a DNA encoding a poiypeptide which has an amino acid sequence derived from the amino acid sequence as shown in SEQ ID NO:
- 2 by substitution, deletion and/or addition of 1-10 amino acid residues and yet has cysteine sulfinic acid decarboxylase activity;
- (c) a DNA encoding a polypeptide having 98% or more amino acid sequence homology with the amino acid sequence as shown in SEQ ID NO:
- 2 and yet having cysteine sulfinic acid decarboxylase activity;
- (d) a DNA having the nucleotide sequence as shown in SEQ ID NO:
- 1;
- (e) a DNA which hybridizes to a DNA complementary to a DNA having the nucleotide sequence as shown in SEQ ID NO:
- 1 under high stringent conditions of 65° C., 2×SSC and 0.1% SDS, and yet encodes a polypeptide having cysteine sultinic acid decarboxylase activity.
A cell which has a transferred DNA encoding cysteine sultinic acid decarboxylase and a transferred DNA encoding a desired antibody.
A cell capable of high-yield production of β-alanine which which has been transformed with a DNA encoding cysteine sulfinic acid decarboxylase.
Dependencies are inferred from the claim text.