EP0155832A2
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Stable proteins, methods for their construction, plasmids and other DNA encoding them, cell cultures harboring such plasmids
Assignee: Genentech Inc
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Filing: 1985-03-15
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Granted: —
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Expires: 2005-03-15
Abstract
Abstract 57 Proteins are mutated in order to introduce disulfide linkages at sites not normally found in the wild type protein. Additional disulfide bonds formed in this fashion stabilize the proteins from inactivation.
Inventors
- 👤 Ronald Burnell Wetzel
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-24 13:04
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/14; C12N9/24; C12N9/2402; C12N9/2462; C12N15/00; C12N15/09; C12N15/10; C12P; C12P21/00; C12P21/02
Claims Analysis
Total Claims: 43
Claim 1
Independent Claim
29 words
An active protein which contains at least one disulfide bond formed between cysteine residues in addition to those which may exist in the naturally occurring form of said protein.
Claim 2
Dependent Claim
29 words
An active protein which contains at least one disulfide bond formed between cysteine residues in addition to those which may exist in the naturally occurring form of said protein.
Claim 3
Dependent Claim
14 words
The protein of claim 1 wherein said protein has no naturally occurring disulfide bonds.
Claim 4
Dependent Claim
13 words
The protein of claim 1 wherein said protein has naturally occurring disulfide bonds.
Claim 5
Dependent Claim
31 words
The protein of any one of the preceding claims wherein the beta carbons of the cysteines forming the additional disulfide bond are separated by a distance of 3.5 to 8 angstroms.
Claim 6
Dependent Claim
38 words
The protein of any one of claims 1, 2 and 3 wherein at least one cysteine residue is introduced to replace a wild type residue, whose alpha carbons are separated by a distance of 3.5 to 10 angstroms.
Claim 7
Dependent Claim
17 words
The protein of any one of the preceding claims wherein said protein retains activity against inactivating forces.
Claim 8
Dependent Claim
26 words
The protein of claim 6 wherein said inactivating forces are selected from heat, pH extremes, presence of denaturing chemicals, oxidation, hydrolysis, temperature variations or degradative enzymes.
Claim 9
Dependent Claim
15 words
The protein of claim 1 wherein said active protein is a hormone or an enzyme.
Claim 10
Dependent Claim
12 words
The protein of claim 8 wherein said protein facilitates lysis of microorganisms.
Claim 11
Dependent Claim
11 words
The protein of claim 9 wherein said protein is a lysozyme.
Claim 12
Dependent Claim
11 words
The protein of claim 10 wherein said protein is T4 lysozyme.
Claim 13
Dependent Claim
17 words
The protein of claim 11 wherein said T4 lysozyme contains cysteine substitution for isoleucine at position three.
Claim 14
Dependent Claim
7 words
DNA encoding the protein of claim 1.
Claim 15
Dependent Claim
8 words
A plasmid containing the DNA of claim 13.
Claim 16
Dependent Claim
22 words
A plasmid having DNA encoding in a protein cysteines which can be oxidized to form at least one non-naturally occurring disulfide bond.
Claim 17
Dependent Claim
22 words
A plasmid having DNA encoding in a protein cysteines which can be oxidized to form at least one non-naturally occurring disulfide bond.
Claim 18
Dependent Claim
57 words
A process for producing an active protein which contains at least one disulfide bond in addition to those which may exist in the naturally occurring form of said protein comprising the introduction of at least one disulfide bond between a naturally occurring cysteine and a cysteine which has been introduced in place of a wild type residue.
Claim 19
Dependent Claim
57 words
A process for producing an active protein which contains at least one disulfide bond in addition to those which may exist in the naturally occurring form of said protein comprising the introduction of at least one disulfide bond between a naturally occurring cysteine and a cysteine which has been introduced in place of a wild type residue.
Claim 20
Dependent Claim
55 words
A process for producing an active protein which contains at least one disulfide bond in addition to those which may exist in the naturally occurring form of said protein comprising the introduction of at least one disulfide bond between two cysteine residues each of which has been introduced in place of a wild type residue.
Claim 21
Dependent Claim
55 words
A process for producing an active protein which contains at least one disulfide bond in addition to those which may exist in the naturally occurring form of said protein comprising the introduction of at least one disulfide bond between two cysteine residues each of which has been introduced in place of a wild type residue.
Claim 22
Dependent Claim
48 words
A process for producing an active protein which contains at least one disulfide bond in addition to those which may exist in the naturally occurring form of said protein comprising the introduction of at least one disulfide bond between two wild type residues both of which are cysteine.
Claim 23
Dependent Claim
48 words
A process for producing an active protein which contains at least one disulfide bond in addition to those which may exist in the naturally occurring form of said protein comprising the introduction of at least one disulfide bond between two wild type residues both of which are cysteine.
Claim 24
Dependent Claim
3 words
The plasmid pT4Lys3CtacII.
Claim 25
Dependent Claim
3 words
The plasmid pT4Lys3CtacII.
Claim 26
Dependent Claim
26 words
A culture comprising cells harboring a plasmid of claim 14 or claim 15 capable of producing a protein of any one of claims 1 to 12.
Claim 27
Dependent Claim
57 words
A process for producing an active protein which contains at least one disulfide bond in addition to those which may exist in the naturally occurring form of said protein comprising the introduction of at least one disulfide bond between a naturally occurring cysteine and a cysteine which has been introduced in place of a wild type residue.
Claim 28
Dependent Claim
57 words
A process for producing an active protein which contains at least one disulfide bond in addition to those which may exist in the naturally occurring form of said protein comprising the introduction of at least one disulfide bond between a naturally occurring cysteine and a cysteine which has been introduced in place of a wild type residue.
Claim 29
Dependent Claim
54 words
A process for producing an active protein which contains-at least one disulfide bond in addition to those which may exist in the naturally occurring form of said protein comprising the introduction of at least one disulfide bond between two cysteine residues each of which has been introduced in place of a wild type residue.
Claim 30
Dependent Claim
54 words
A process for producing an active protein which contains-at least one disulfide bond in addition to those which may exist in the naturally occurring form of said protein comprising the introduction of at least one disulfide bond between two cysteine residues each of which has been introduced in place of a wild type residue.
Claim 31
Dependent Claim
48 words
A process for producing an active protein which contains at least one disulfide bond in addition to those which may exist in the naturally occurring form of said protein comprising the introduction of at least one disulfide bond between two wild type residues both of which are cysteine.
Claim 32
Dependent Claim
48 words
A process for producing an active protein which contains at least one disulfide bond in addition to those which may exist in the naturally occurring form of said protein comprising the introduction of at least one disulfide bond between two wild type residues both of which are cysteine.
Claim 33
Dependent Claim
20 words
The process of any one of claims 1, 2 and 3 wherein said protein has no naturally occurring disulfide bonds.
Claim 34
Dependent Claim
19 words
The process of any one of claims 1, 2 and 3 wherein said protein has naturally occurring disulfide bonds.
Claim 35
Dependent Claim
31 words
The process of any one of the preceding claims wherein the beta carbons of the cysteines forming the additional disulfide bond are separated by a distance of 3.5 to 8 angstroms.
Claim 36
Dependent Claim
37 words
The process of any one of claims 1 to 5 wherein at least one cysteine residue is introduced to replace a wild type residue, whose alpha carbons are separated by a distance of 3.5 to 10 angstroms.
Claim 37
Dependent Claim
17 words
The process of any one of the preceding claims wherein said protein retains activity against inactivating forces.
Claim 38
Dependent Claim
26 words
The process of claim 8 wherein said inactivating forces are selected from heat, pH extremes, presence of denaturing chemicals, oxidation, hydrolysis, temperature variations or degradative enzymes.
Claim 39
Dependent Claim
21 words
The process of any one of claims 1, 2 and 3 wherein said active protein is a hormone or an enzyme.
Claim 40
Dependent Claim
12 words
The process of claim 10 wherein said protein facilitates lysis of microorganisms.
Claim 41
Dependent Claim
11 words
The protein of claim 11 wherein said protein is a lysozyme.
Claim 42
Dependent Claim
11 words
The protein of claim 12 wherein said protein is T4 lysozyme.
Claim 43
Dependent Claim
17 words
The process of claim 13 wherein said T4 lysozyme contains cysteine substitution for isoleucine at position three.
References Cited (Backward Citations)
No citation records indexed.
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