US6294353B1
Expired / Lapsed
US
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Targeted hetero-association of recombinant proteins to multi-functional complexes
Assignee: Morphosys GmbH
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Filing: 1995-10-20
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Granted: 2001-09-25
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Expires: 2015-10-20
Abstract
Abstract The present invention relates to a method for targeted assembly of distinct active peptide or protein domains into a single complex and to such complexes. The invention relates particularly to the fusion of peptide or protein domains to complementary association domains which are derived from a single tertiary or quaternary structure by segmentation. The association domains are designed to assemble in a complementary fashion, thereby providing multifunctional (poly)peptides.
Inventors
- 👤 Peter Pack
- 👤 Andrei Lupas
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 15:34
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/001; A; A61; A61P; A61P7/00; A61P7/02; C07K16/00; C07K16/46; C07K16/468; A61K; A61K38/00
Claims Analysis
Total Claims: 30
Claim 1
Independent Claim
61 words
A multifunctional polypeptide comprising from amino-terminal to carboxy-terminal: (a) a first functional domain; (b) an assembly device consisting of a segment of a hydrophobic core of a parent polypeptide, said assembly device capable of forming heteromultimers, said assembly device not naturally occurring with the first functional domain; (c) a second functional domain which is not naturally associated with the assembly device.
Claim 2
Dependent Claim
61 words
A multifunctional polypeptide comprising from amino-terminal to carboxy-terminal: (a) a first functional domain; (b) an assembly device consisting of a segment of a hydrophobic core of a parent polypeptide, said assembly device capable of forming heteromultimers, said assembly device not naturally occurring with the first functional domain; (c) a second functional domain which is not naturally associated with the assembly device.
Claim 3
Dependent Claim
19 words
The multifunctional polypeptide according to claim 1 , wherein said parent polypeptide is ubiquitin, acyl-phosphatase, IL2, calbindin or apomyoglobin.
Claim 4
Dependent Claim
20 words
The multifunctional polypeptide according to claim 1 or 2 , wherein said hydrophobic core of parent polypeptide is biologically active.
Claim 5
Dependent Claim
27 words
The multifunctional polypeptide according to claim 1 , wherein at least one of said functional domains is a fragment derived from a member of the immunoglobulin superfamily.
Claim 6
Dependent Claim
23 words
The multifunctional polypeptide according to claim 4 , wherein two of said functional domains are fragments derived from members of the immunoglobulin superfamily.
Claim 7
Dependent Claim
16 words
The multifunctional polypeptide according to claim 4 or 5 , wherein said fragments are antibody fragments.
Claim 8
Dependent Claim
37 words
The multifunctional polypeptide according to claim 1 , wherein at least one of said functional domains is a biologically active molecule or a derivative thereof other than a fragment derived from a member of the immunoglobulin superfamily.
Claim 9
Dependent Claim
20 words
The multifunctional polypeptide according to claim 1 , wherein folding of the assembly device is stabilised by a covalent bond.
Claim 10
Dependent Claim
27 words
The multifunctional polypeptide according to claim 1 , wherein at least one of said functional domains is coupled to said assembly device via a flexible peptide linker.
Claim 11
Dependent Claim
18 words
The multifunctional polypeptide according to claim 9 , wherein said flexible peptide linker is an antibody hinge region.
Claim 12
Dependent Claim
75 words
The multifunctional polypeptide according to claim 1 , wherein at least one of said functional domains is an enzyme, a toxin, a cytokine, a metal binding site, a metal binding protein, a soluble receptor, a DNA-binding domain, a transcription factor, an immunoglobulin, a bioactive peptide of 5 to 15 amino acid residues, a peptide hormone, a growth factor, a lectin, a lipoprotein, or a peptide which is able to bind to an independent binding entity.
Claim 13
Dependent Claim
31 words
A composition comprising at least two multifunction polypetides of claim 1 , wherein the multifunctional polypeptides form multimeric multifunctional polypeptides through the interaction of the assembly devices between the multifunctional polypeptides.
Claim 14
Dependent Claim
22 words
The composition according to claim 12 , wherein at least one of said assembly devices or functional domains is of human origin.
Claim 15
Dependent Claim
18 words
A pharmaceutical composition comprising the composition according to claim 12 optionally in combination with a pharmaceutically acceptable carrier.
Claim 16
Dependent Claim
12 words
A diagnostic composition comprising the multifunctional polypeptide according to claim 1 .
Claim 17
Dependent Claim
11 words
A DNA sequence encoding the multifunctional polypeptide of claim 1 .
Claim 18
Dependent Claim
12 words
A vector comprising at least one DNA sequence of claim 16 .
Claim 19
Dependent Claim
11 words
The vector of claim 17 , which is a bicistronic vector.
Claim 20
Dependent Claim
14 words
A host cell transformed with at least one vector according to claim 17 .
Claim 21
Dependent Claim
21 words
The host cell according to claim 19 , wherein said host cell is a mammalian, yeast, insect, plant or bacterial cell.
Claim 22
Dependent Claim
41 words
A vector cassette characterised in that it comprises a DNA sequence encoding an assembly device comprised in the multifunctional polypeptide of claim 1 , and additionally at least one cloning site for inserting a DNA sequence encoding a further functional domain.
Claim 23
Dependent Claim
74 words
A vector cassette characterised in that it comprises a DNA sequence encoding the assembly device according to claim 1 and suitable restriction sites for the cloning of DNA sequences encoding the functional domains according to claim 1 , such that, after the insertion of the DNA sequences encoding the functional domains into said restriction sites, expression from the vector cassette in a suitable host results in the multifunctional polypeptide according to claim 1 .
Claim 24
Dependent Claim
15 words
A kit comprising at least one vector cassette according to claim 21 or 22 .
Claim 25
Dependent Claim
22 words
The vector cassette according to claim 20 or 22 characterised in that it comprises the inserted DNA sequence(s) encoding said function domain(s).
Claim 26
Dependent Claim
15 words
A host cell transformed with at least one vector cassette according to claim 24 .
Claim 27
Dependent Claim
21 words
The host cell according to claim 25 , wherein said host cell is a mammalian, yeast, insect, plant or bacterial cell.
Claim 28
Dependent Claim
44 words
A method for the production of a multifunctional polypeptide according to claim 1 , which comprises culturing the host cell according to claim 19 , 20 , 25 or 26 in a suitable medium, and recovering said multifunctional polypeptide produced by said host cell.
Claim 29
Dependent Claim
75 words
A method for the production of a composition according to claim 12 which comprises culturing at least two host cells according to claim 20 , 25 or 26 in a suitable medium, said host cells each producing only one assembly device attached to at least one further functional domain, recovering the multifunctional polypeptides, mixing thereof under mildly denaturing conditions and allowing in vitro interaction of the multifunctional polypeptide assembly devices according to claim 12 .
Claim 30
Dependent Claim
60 words
A method for the production of a composition of claim 12 which comprises the steps of (a) culturing a host cell according to claim 19 , 20 , 25 or 26 , wherein said host cell is capable of producing at least two different multifunctional polypeptides according to claim 1 ; and (b) allowing multimerization of multifunctional polypeptide assembly devices.
References Cited (Backward Citations)
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 2004-09-24 | 2005-03-26 | 2005-09-25 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2008-09-24 | 2009-03-26 | 2009-09-25 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2012-09-24 | 2013-03-26 | 2013-09-25 | $7700.00 | Window Closed / Lapsed |