Method for producing protein by precipitation
AssigneeAjinomoto Co., Inc.
InventorsTakahiro Nonaka, Teruhisa Mannen, Noriko Tsurui
Life of this patent
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Abstract
A method of purifying a fusion protein, including: (1) adjusting the pH of an aqueous phase, containing a fusion protein which is a fusion of a protein having a self-assembly capability and a tamet protein, and which aqueous phase has a first pH, to a second pH, to obtain a remaining aqueous phase and a solid fraction containing an amount of the fusion protein; (2) separating the solid fraction from the remaining aqueous phase, to obtain a separated solid fraction; and (3) dissolving the separated solid fraction in a solution having a pH of 12 or lower but higher than the second pH by at least 0.1 pH units, wherein the protein having a self-assembly capability is a cell surface protein.
Claims33
A method of purifying a fusion protein, comprising:
- (1) adjusting the pH of
- an aqueous phase, comprising a fusion protein which is a fusion of a protein having a self-assembly capability and a target protein, and which aqueous phase has a first pH, to a second pH, to obtain a remaining aqueous phase and a solid fraction comprising an amount of said fusion protein;
- (2) separating said solid fraction from said remaining aqueous phase, to obtain a separated solid fraction;
- and
- (3) dissolving said separated solid fraction in a solution having a pH of 12 or lower but higher than said second pH by at least 0.1 pH units,
- wherein said protein having a self-assembly capability is a cell surface protein that is a CspB mature protein or a portion thereof;
- and
- wherein (i) said target protein has 10 to 300 amino acid residues, or (ii) the ratio of the number of amino acid residues in the target protein to the number of amino acid residues in the CspB mature protein or portion thereof is in a range of 0.3 to 14.
A method for producing a target protein, comprising:
- (1) adjusting the pH of
- an aqueous phase, comprising a fusion protein which is a fusion of a protein having a self-assembly capability, a target protein, and an amino acid sequence capable of enzymatic cleavage or a chemical cleavage located between said protein haying a self-assembly capability and said target protein, and which aqueous phase has a first pH, to a second pH, to obtain a remaining aqueous phase and a solid fraction comprising an amount of said fusion protein;
- (2) separating said solid fraction from said remaining aqueous phase, to obtain a separated solid fraction;
- (3) dissolving said separated solid fraction in a solution having a pH of 12 or lower but higher than said second pH by at least 0.1 pH units;
- and
- (4) enzymatically or chemically cleaving said fusion protein at said site of said amino acid sequence located between said protein having a self-assembly capability and said target protein, either simultaneously with, during, or after said dissolving (3), to obtain said target protein,
- wherein said protein having a self-assembly capability is a cell surface protein that is a CspB mature protein or a portion thereof;
- and
- wherein (i) said target protein has 10 to 300 amino acid residues, or (ii) the ratio of the number of amino acid residues in the target protein to the number of amino acid residues in the CspB mature protein or portion thereof is in a range of 0.3 to 14.
A method for isolating a solid of a fusion protein, comprising:
- (1) adjusting the pH of an aqueous phase, comprising a fusion protein which is a fusion of a protein having a self-assembly capability and a target protein, and which aqueous phase has a first pH, to a second pH, to obtain a remaining aqueous phase and a solid fraction comprising an amount of said fusion protein;
- and
- (2) separating said solid fraction from said remaining aqueous phase, to obtain a separated solid fraction which comprises said fusion protein,
- wherein said protein having a self-assembly capability is a cell surface protein that is a CspB mature protein or a portion thereof;
- and
- wherein (i) said target protein has 10 to 300 amino acid residues, or (ii) the ratio of the number of amino acid residues in the target protein to the number of amino acid residues in the CspB mature protein or portion thereof is in a range of 0.3 to 14.
Dependencies are inferred from the claim text.