Method for preparing a physiologically active il-18 polypeptide
AssigneeSmithKline Beecham Corporation
InventorsKyung O Johanson, Robert B Kirkpatrick, Allan R Shatzman, Yen Sen Ho, Patrick McDevitt
Life of this patent
Estimating the term…
Abstract
A method for producing a physiologically active polypeptide, comprising contacting a precursor polypeptide with an activating enzyme, or co-expressing the polypeptide with an activating protease.
Claims24
A method for producing an active polypeptide from a precursor polypeptide polypeptide having a protease activation motif comprising the amino acid sequence XEYD, wherein X is selected from a group of amino acids consisting of W, L, F, Y, I, V, D, or E;
- and Y is selected from a group of amino acids consisting of H, I, A, T, S, P or E, comprising:
- (i) contacting caspase 4 with the precursor polypeptide and
- (ii) purifying the active polypeptide.
A method for producing an active polypeptide from a precursor polypeptide polypeptide having a protease activation motif comprising the amino acid sequence XEYD, wherein X is selected from a group of amino acids consisting of W, L, F, Y, I, V, D, or E;
- and Y is selected from a group of amino acids consisting of H, I, A, T, S, P or E, comprising:
- (i) contacting caspase 5 with the precursor polypeptide and
- (ii) purifying the active polypeptide.
A method for producing an active IL-18 polypeptide from a precursor IL-18 polypeptide, comprising:
- (i) contacting caspase 4 with a precursor IL-18 polypeptide;
- and
- (ii) purifying the active IL-18 polypeptide.
A method for producing an active IL-18 polypeptide from a precursor IL-18 polypeptide, comprising:
- (i) contacting caspase 5 with the precursor IL-18 polypeptide;
- and
- (ii) purifying the active IL-18 polypeptide.
A method for producing an active human IL-18 polypeptide comprising:
- (i) contacting caspase 4 with a human precursor IL-18 polypeptide to convert said precursor polypeptide into the active human IL-18 polypeptide, said precursor IL-18 polypeptide comprising the amino acid sequence of SEQ ID NO:
- 1, or a portion thereof, which maintains the aspartic acid 36 and tyrosine 37 residues in the amino sequence of SEQ ID NO:
- 1 as an N-terminal sequence;
- and
- (ii) purifying the active human IL-18 polypeptide.
A method for producing an active human IL-18 polypeptide comprising:
- (i) contacting caspase 5 with a human precursor IL-18 polypeptide to convert said precursor polypeptide into the active human IL-18 polypeptide, said precursor IL-18 polypeptide comprising the amino acid sequence of SEQ ID NO:
- 1, or a portion thereof, which maintains the aspartic acid 36 and tyrosine 37 residues in the amino sequence of SEQ ID NO:
- 1 as an N-terminal sequence;
- and
- (ii) purifying the active IL-18 polypeptide.
A method for producing an active polypeptide from a precursor polypeptide, comprising:
- (i) co-expressing bicistronically caspase 4 with the precursor polypeptide;
- and
- (ii) purifying the active polypeptide.
A method for producing an active polypeptide from a precursor polypeptide, comprising:
- (i) co-expressing bicistronically caspase 5 with the precursor polypeptide;
- and
- (ii) purifying the active polypeptide.
A method for producing active IL-18 polypeptide from a precursor IL-18 polypeptide, comprising:
- (i) co-expressing bicistronically ubiquitinase with a ubiquitin/IL-18 fusion;
- and
- (ii) purifying the active IL-18 polypeptide.
A method for producing an active IL-18 polypeptide from a precursor polypeptide, comprising:
- (i) co-expressing bicistronically caspase 4 with the precursor IL-18 polypeptide;
- and
- (ii) purifying the active IL-18 polypeptide.
A method for producing an active IL-18 polypeptide from a precursor IL-18 polypeptide, comprising:
- (i) co-expressing bicistronically caspase 5 with the precursor IL-18 polypeptide;
- and
- (ii) purifying the active IL-18 polypeptide.
A method for producing active human IL-18 polypeptide from precursor human IL-18 polypeptide, comprising:
- (i) co-expressing bicistronically ubiquitinase with a ubiquitin/IL-18 having the amino acid sequence of SEQ ID NO:10;
- and
- (ii) purifying the active human IL-18 polypeptide.
A method for producing an active human IL-18 polypeptide from a human precursor IL-18 polypeptide, comprising:
- (i) co-expressing bicistronically caspase 4, having the amino acid sequence of SEQ ID NO:4, with the human precursor IL-18 polypeptide;
- and
- (ii) purifying the active IL-18 polypeptide.
A method for producing an active human IL-18 polypeptide from a human precursor IL-18 polypeptide, comprising:
- (i) co-expressing bicistronically caspase 5, having the amino acid sequence of SEQ ID NO:6, with the human precursor IL-18 polypeptide;
- and
- (ii) purifying the active IL-18 polypeptide.
Dependencies are inferred from the claim text.