US6107059A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Peptide library and screening method
Assignee: Affymax Technologies NV
•
Filing: 1992-04-29
•
Granted: 2000-08-22
•
Expires: 2012-04-29
Abstract
Abstract A random peptide library constructed by transforming host cells with a collection of recombinant vectors that encode a fusion protein comprised of a carrier protein fused to a random peptide through a proteolytic cleavage site can be used to identify ligands that bind to a receptor. The screening method results in the formation of a complex comprising the fusion protein bound to a receptor through the random peptide ligand, and the random peptide can easily be identified and analyzed by virtue of the carrier protein and associated proteolytic cleavage site.
Inventors
- 👤 Charles P. Hart
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 06:08
CPC Classifications
C; C40; C40B; C40B40/00; C40B40/02; C07; C07K; C07K1/00; C07K1/04; C07K1/047; C07K14/00; C07K14/37; C07K14/39; C07K14/395; C07K14/435; C07K14/665; C12; C12N; C12N15/00; C12N15/09; C12N15/10; C12N15/1034; C12N15/1037; C12N15/11; C12N15/62; C07K2319/00; C07K2319/20; C07K2319/24; C07K2319/70; C07K2319/74; C07K2319/75; C07K2319/80; C07K2319/81
Claims Analysis
Total Claims: 9
Claim 1
Independent Claim
100 words
A method of generating a peptide library comprising the steps of: (a) transforming host cells with a set of recombinant DNA vectors, each member of which encodes a different tripartite fusion protein consisting essentially of a carrier protein, a peptide, and a proteolytic cleavage site between said carrier protein and said peptide, such that at least 1000 and up to about 10 8 different transformants are formed, wherein said different members differ from one another with respect to the peptide encoded; and (b) culturing said host cells transformed in step (a) under conditions suitable for expression of the fusion protein.
Claim 2
Independent Claim
100 words
A method of generating a peptide library comprising the steps of: (a) transforming host cells with a set of recombinant DNA vectors, each member of which encodes a different tripartite fusion protein consisting essentially of a carrier protein, a peptide, and a proteolytic cleavage site between said carrier protein and said peptide, such that at least 1000 and up to about 10 8 different transformants are formed, wherein said different members differ from one another with respect to the peptide encoded; and (b) culturing said host cells transformed in step (a) under conditions suitable for expression of the fusion protein.
Claim 3
Dependent Claim
74 words
The method of claim 1, wherein said host cell is E. coli, said vector is a plasmid, said carrier protein is selected from the group consisting of maltose binding protein, streptavidin, His n where n is at least 3, ubiquitin, cellulose binding protein, and glutathione-S-transferase, and said proteolytic cleavage site is selected from the group consisting of a site for Factor Xa cleavage, a site for enterokinase cleavage, and a site for collagenase cleavage.
Claim 4
Dependent Claim
26 words
The method of claim 2, wherein said carrier protein is the maltose binding protein and said proteolytic cleavage site is a site for Factor Xa cleavage.
Claim 5
Dependent Claim
28 words
The method of claim 3, wherein said plasmid further encodes a signal peptide that directs secretion of the tripartite fusion protein into the periplasm of said host cell.
Claim 6
Dependent Claim
17 words
The method of claim 3, wherein said peptide is five to seventy-five amino acid residues in length.
Claim 7
Dependent Claim
19 words
The method of claim 5, wherein said peptide comprises amino acid residues that introduce conformational constraints on the peptide.
Claim 8
Dependent Claim
37 words
The method of claim 6, wherein said peptide assumes a conformation characteristic of a conformation selected from the group consisting of a zinc finger, leucine zipper, cysteine crosslinking via disulphide bridge formation, beta turn, and alpha helix.
Claim 9
Dependent Claim
29 words
The method of claim 1, wherein said peptide library is a secondary library, and said peptide of said tripartite fusion protein is a variation of a lead peptide sequence.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 2003-08-22 | 2004-02-21 | 2004-08-22 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2007-08-22 | 2008-02-21 | 2008-08-22 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2011-08-22 | 2012-02-21 | 2012-08-22 | $7700.00 | Window Closed / Lapsed |