US5403484A
Expired / Lapsed
US
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Viruses expressing chimeric binding proteins
Assignee: Dyax Corp
•
Filing: 1993-01-26
•
Granted: 1995-04-04
•
Expires: 2013-01-26
Abstract
Abstract In order to obtain a novel binding protein against a chosen target, DNA molecules, each encoding a protein comprising one of a family of similar potential binding domains and a structural signal calling for the display of the protein on the outer surface of a chosen bacterial cell, bacterial spore or phage (genetic package) are introduced into a genetic package. The protein is expressed and the potential binding domain is displayed on the outer surface of the package. The cells or viruses bearing the binding domains which recognize the target molecule are isolated and amplified. The successful binding domains are then characterized. One or more of these successful binding domains is used as a model for the design of a new family of potential binding domains, and the process is repeated until a novel binding domain having a desired affinity for the target molecule is obtained. In one embodiment, the first family of potential binding domains is related to bovine pancreatic trypsin inhibitor, the genetic package is M13 phage, and the protein includes the outer surface transport signal of the M13 gene III protein.
Inventors
- 👤 Robert C. Ladner
- 👤 Sonia K. Guterman
- 👤 Bruce L. Roberts
- 👤 William Markland
- 👤 Arthur C. Ley
- 👤 Rachel B. Kent
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 13:36
CPC Classifications
C; C40; C40B; C40B40/00; C40B40/02; C07; C07K; C07K1/00; C07K1/04; C07K1/047; C07K1/107; C07K14/00; C07K14/435; C07K14/43504; C07K14/43513; C07K14/43522; C07K14/795; C07K14/805; C07K14/81; C07K14/8107; C07K14/811; C07K14/8114; C07K14/8117; C12; C12N; C12N15/00; C12N15/09; C12N15/10; C12N15/1034; C12N15/1037; C12N7/00; A; A61; A61K; A61K38/00; C12N2795/00; C12N2795/00011; C12N2795/14011; C12N2795/14111; C12N2795/14122; C12N2795/14141; C12N2795/14143
Claims Analysis
Total Claims: 53
Claim 1
Independent Claim
155 words
A virus bearing on its outer surface a chimeric binding protein, said protein comprising (i) a proteinaceous binding domain, other than a single chain antibody, which is sufficiently stable in structure to have a melting point of at least 40° C., and which binds to a target:, other than the variable domain of an antibody, sufficiently strongly so that the dissociation constant of the binding domain: target complex is less than 10 -6 moles/liter, and (ii) at least a functional portion of a coat protein of said virus, said portion acting, when the chimeric protein is produced in a suitable host cell, to cause the display of the chimeric binding protein or a processed form thereof on the outer surface of the virus, said binding domain being capable of binding to a target material which said coat protein does not preferentially bind, said binding domain being foreign to the native coat proteins of said virus.
Claim 2
Dependent Claim
155 words
A virus bearing on its outer surface a chimeric binding protein, said protein comprising (i) a proteinaceous binding domain, other than a single chain antibody, which is sufficiently stable in structure to have a melting point of at least 40° C., and which binds to a target:, other than the variable domain of an antibody, sufficiently strongly so that the dissociation constant of the binding domain: target complex is less than 10 -6 moles/liter, and (ii) at least a functional portion of a coat protein of said virus, said portion acting, when the chimeric protein is produced in a suitable host cell, to cause the display of the chimeric binding protein or a processed form thereof on the outer surface of the virus, said binding domain being capable of binding to a target material which said coat protein does not preferentially bind, said binding domain being foreign to the native coat proteins of said virus.
Claim 3
Dependent Claim
11 words
The virus of claim 1 wherein the virus is a phage.
Claim 4
Dependent Claim
12 words
The virus of claim 2 wherein the phage is a filamentous phage.
Claim 5
Dependent Claim
13 words
The virus of claim 3 wherein the coat protein is the gIII protein.
Claim 6
Dependent Claim
13 words
The virus of claim 3 wherein the coat protein is the gVIII protein.
Claim 7
Dependent Claim
19 words
The virus of claim 1 wherein the proteinaceous binding domain has a melting point of at least 50° C.
Claim 8
Dependent Claim
15 words
The virus of claim 1 wherein the proteinaceous binding domain features at least two disulfides.
Claim 9
Dependent Claim
28 words
The virus of claim 1 wherein the proteinaceous binding domain has a single disulfide bond and the span of the bond is not more than nine amino acids.
Claim 10
Dependent Claim
21 words
The virus of claim 1 wherein the proteinaceous binding domain contains no more than 30 residues and at least 2 disulfides.
Claim 11
Dependent Claim
21 words
The virus of claim 1 wherein the proteinaceous binding domain contains no more than 60 residues and at least 3 disulfides.
Claim 12
Dependent Claim
21 words
The virus of claim 1 wherein the proteinaceous binding domain contains no more than 80 residues and at least 4 disulfides.
Claim 13
Dependent Claim
14 words
The virus of claim 1 wherein the proteinaceous binding domain is a protease inhibitor.
Claim 14
Dependent Claim
15 words
The virus of claim 12 wherein the proteinaceous binding domain is a serine protease inhibitor.
Claim 15
Dependent Claim
17 words
The virus of claim 1 wherein the proteinaceous binding domain is at least 50% homologous with BPTI.
Claim 16
Dependent Claim
34 words
The virus of claim 1 wherein the chimeric protein further comprises a linker peptide, linking said binding domain to said coat protein or functional portion thereof, which is specifically cleavable by a site-specific protease.
Claim 17
Dependent Claim
21 words
The virus of claim 15 wherein the site-specific protease is Factor Xa, Factor XIa, Kallikvein, thrombin, Factor XIIa, collagenase or enterokinase.
Claim 18
Dependent Claim
18 words
A library of virus according to claim 1, said library collectively displaying a plurality of different binding domains.
Claim 19
Dependent Claim
16 words
The library of claim 17 wherein at least 10 4 different proteinaceous binding domains are displayed.
Claim 20
Dependent Claim
34 words
The library of claim 17, wherein at least some of the displayed binding domains have amino acid sequences which are not identical to an amino acid sequence encoded by a naturally occurring DNA sequence.
Claim 21
Dependent Claim
31 words
The virus of claim 1 wherein the proteinaceous binding domain has an amino acid sequence which is not identical to an amino acid sequence encoded by a naturally occurring DNA sequence.
Claim 22
Dependent Claim
34 words
The virus of claim 1 wherein the amino acid sequence differences among the binding domains of said library are limited to no more than about 20 corresponding amino acid residue positions of said domains.
Claim 23
Dependent Claim
78 words
The virus of claim 1 wherein the binding domain is at least substantially homologous with a binding domain selected from the group consisting of the binding domains of bovine pancreatic trypsin inhibitor, crambin, Cucurbita maxima trypsin inhibitor III, heat stable enterotoxin of Escherichia coli, α Conotoxin GI, λ Conotroxin GIII, ω Conotoxin GIV, apamin, charybdtoxin, secretory leukocyte protease inhibitor, cystatin, eglin, barley protease inhibitor, ovomucoid, T4 lysozyme, hen egg white lysozyme, ribonuclease, azurin, tumor necrosis factor, and CD4.
Claim 24
Dependent Claim
21 words
The virus of claim 1, said virus further bearing on its outer surface the corresponding wild-type coat protein of said virus.
Claim 25
Dependent Claim
8 words
A library of virus according to claim 23.
Claim 26
Dependent Claim
22 words
The virus of claim 1 wherein the proteinaceous binding domain is coupled essentially to the amino terminal of the mature coat protein.
Claim 27
Dependent Claim
146 words
A chimeric binding protein comprising (i) a proteinaceous binding domain, other than a single chain antibody, which is sufficiently stable in structure to have a melting point of at least 40° C., and which binds to a target, other than the variable domain of an antibody, sufficiently strongly so that the disassociation constant of the binding domain: target complex is less that 10 -6 moles/liter, and (ii) at least a functional portion of a coat protein of a virus, said portion acting, when the chimeric protein is produced in a suitable host cell, to cause the display of the chimeric binding protein or a processed form thereof on the outer surface of the virus, said binding domain being capable of binding to a target material which said coat protein does not preferentially bind, said binding domain being foreign to the native coat proteins of said virus.
Claim 28
Dependent Claim
146 words
A chimeric binding protein comprising (i) a proteinaceous binding domain, other than a single chain antibody, which is sufficiently stable in structure to have a melting point of at least 40° C., and which binds to a target, other than the variable domain of an antibody, sufficiently strongly so that the disassociation constant of the binding domain: target complex is less that 10 -6 moles/liter, and (ii) at least a functional portion of a coat protein of a virus, said portion acting, when the chimeric protein is produced in a suitable host cell, to cause the display of the chimeric binding protein or a processed form thereof on the outer surface of the virus, said binding domain being capable of binding to a target material which said coat protein does not preferentially bind, said binding domain being foreign to the native coat proteins of said virus.
Claim 29
Dependent Claim
11 words
The protein of claim 26 wherein the virus is a phage.
Claim 30
Dependent Claim
12 words
The protein of claim 27 wherein the phage is a filamentous phage.
Claim 31
Dependent Claim
13 words
The protein of claim 28 wherein the coat protein is the gIII protein.
Claim 32
Dependent Claim
13 words
The protein of claim 28 wherein the coat protein is the gVIII protein.
Claim 33
Dependent Claim
18 words
The protein of claim 26 wherein the binding domain has a melting point of at least 50° C.
Claim 34
Dependent Claim
14 words
The protein of claim 26 wherein the binding domain features at least two disulfides.
Claim 35
Dependent Claim
28 words
The protein of claim 26 wherein the proteinaceous binding domain has a single disulfide bond and the span of the bond is not more than nine amino acids.
Claim 36
Dependent Claim
21 words
The protein of claim 26 wherein the proteinaceous binding domain contains no more than 30 residues and at least 2 disulfides.
Claim 37
Dependent Claim
21 words
The protein of claim 26 wherein the proteinaceous binding domain contains no more than 60 residues and at least 3 disulfides.
Claim 38
Dependent Claim
21 words
The protein of claim 28 wherein the proteinaceous binding domain contains no more than 80 residues and at least 4 disulfides.
Claim 39
Dependent Claim
14 words
The protein of claim 26 wherein the proteinaceous binding domain is a protease inhibitor.
Claim 40
Dependent Claim
15 words
The protein of claim 37 wherein the proteinaceous binding domain is a serine protease inhibitor.
Claim 41
Dependent Claim
17 words
The protein of claim 26 wherein the proteinaceous binding domain is at least 50% homologous with BPTI.
Claim 42
Dependent Claim
35 words
The protein of claim 26 wherein the chimeric binding protein further comprises a linker peptide, linking said binding domain to said coat protein or functional portion thereof, which is specifically cleavable by a site-specific protease.
Claim 43
Dependent Claim
21 words
The protein of claim 40 wherein the site-specific protease is Factor Xa, Factor XIa, Kallikvein, thrombin, Factor XIIa, collagenase or enterokinase.
Claim 44
Dependent Claim
31 words
The protein of claim 26 wherein the proteinaceous binding domain has an amino acid sequence which is not identical to an amino acid sequence encoded by a naturally occurring DNA sequence.
Claim 45
Dependent Claim
22 words
The protein of claim 26 wherein the proteinaceous binding domain is coupled essentially to the amino terminal of the mature coat protein.
Claim 46
Dependent Claim
77 words
A fusion protein comprising (a) a carrier protein moiety essentially corresponding to a mature gene III protein of a filamentous phage, said carrier protein moiety acting, when the fusion protein is produced in a suitable host cell infected by the phage, to cause the display of the fusion protein or a processed form thereof on the surface of the phage, and (b) a foreign peptide or protein coupled to the amino terminal of said carrier protein moiety.
Claim 47
Dependent Claim
77 words
A fusion protein comprising (a) a carrier protein moiety essentially corresponding to a mature gene III protein of a filamentous phage, said carrier protein moiety acting, when the fusion protein is produced in a suitable host cell infected by the phage, to cause the display of the fusion protein or a processed form thereof on the surface of the phage, and (b) a foreign peptide or protein coupled to the amino terminal of said carrier protein moiety.
Claim 48
Dependent Claim
64 words
A recombinant filamentous phage bearing a fusion protein according to claim 44, upon its outer surface, said carrier protein moiety being integrated into the coat of the phage, said foreign peptide or protein being capable of binding specifically to a target which said phage does not specifically bind and being of an amino acid sequence foreign to the coat proteins native to said phage.
Claim 49
Dependent Claim
20 words
A library of recombinant phage according to claim 45, said library displaying a plurality of different foreign peptides or proteins.
Claim 50
Dependent Claim
75 words
A fusion protein comprising (a) at least a functional portion of a mature gene VIII protein of a filamentous phage, said portion acting, when the fusion protein is produced in a suitable host cell infected by the phage to cause the display of the fusion protein or a processed form thereof on the surface of the phage, and (b) a foreign peptide or protein coupled to said functional portion of said mature gene VIII protein.
Claim 51
Dependent Claim
75 words
A fusion protein comprising (a) at least a functional portion of a mature gene VIII protein of a filamentous phage, said portion acting, when the fusion protein is produced in a suitable host cell infected by the phage to cause the display of the fusion protein or a processed form thereof on the surface of the phage, and (b) a foreign peptide or protein coupled to said functional portion of said mature gene VIII protein.
Claim 52
Dependent Claim
68 words
A recombinant filamentous phage bearing a fusion protein according to claim 47, upon its outer surface, said functional portion of the gene VIII protein being integrated into the coat of the phage, said foreign peptide or protein being capable of binding specifically to a target which said phage does not specifically bind and being of an amino acid sequence foreign to the coat proteins native to said phage.
Claim 53
Dependent Claim
20 words
A library of recombinant phage according to claim 48, said library displaying a plurality of different foreign peptides or proteins.
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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) | 1998-04-03 | 1998-10-03 | 1999-04-04 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2002-04-03 | 2002-10-03 | 2003-04-04 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2006-04-03 | 2006-10-03 | 2007-04-04 | $7700.00 | Window Closed / Lapsed |