Patent US4752585A successfully refreshed from Google Patents.
US4752585A
Expired / Lapsed
US
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Oxidation-resistant muteins
Assignee: Novartis Vaccines and Diagnostics Inc
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Filing: 1986-08-05
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Granted: 1988-06-21
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Expires: 2006-08-05
Abstract
Abstract A biologically active reference therapeutic protein is protected against oxidation by a method involving substituting a conservative amino acid for each methionyl residue susceptible to chloramine T or peroxide oxidation, wherein additional, non-susceptible methionyl residues are not so substituted. The oxidation-resistant mutein so produced is preferably a human mutein of interleukin-2 or interferon-β, and the conservative amino acid is most preferably alanine.
Inventors
- 👤 Kirston E. Koths
- 👤 Robert F. Halenbeck
- 👤 Michael A. Innis
- 👤 Alice M. Wang
- 👤 Ze'ev Shaked
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-04 18:37
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/52; C07K14/54; C07K14/55; Y; Y10; Y10S; Y10S930/00; Y10S930/01; Y10S930/14; Y10S930/141; Y10S930/142
Claims Analysis
Total Claims: 14
Claim 1
Independent Claim
117 words
A recombinant DNA sequence that encodes an interleukin-2 (IL-2) mutein having therapeutically useful biological activity in comparison with a reference protein, wherein said reference protein is either mature native recombinant IL-2 having the primary amino acid sequence shown in FIG. 1 or the des ala 1 ser 125 IL-2 mutein thereof, and facilitating the production of a less heterogeneous protein preparation, as determined by reverse phase high performance liquid chromatography analysis, in comparison with said reference protein, wherein said IL-2 mutein is oxidation-resistant where each methionine residue of said reference protein that is susceptible to chloramine T or hydrogen peroxide oxidation is replaced by a conservative amino acid, and having additional, non-susceptible methionine residues not so substituted.
Claim 2
Dependent Claim
117 words
A recombinant DNA sequence that encodes an interleukin-2 (IL-2) mutein having therapeutically useful biological activity in comparison with a reference protein, wherein said reference protein is either mature native recombinant IL-2 having the primary amino acid sequence shown in FIG. 1 or the des ala 1 ser 125 IL-2 mutein thereof, and facilitating the production of a less heterogeneous protein preparation, as determined by reverse phase high performance liquid chromatography analysis, in comparison with said reference protein, wherein said IL-2 mutein is oxidation-resistant where each methionine residue of said reference protein that is susceptible to chloramine T or hydrogen peroxide oxidation is replaced by a conservative amino acid, and having additional, non-susceptible methionine residues not so substituted.
Claim 3
Dependent Claim
21 words
The recombinant DNA sequence of claim 1 wherein the reference protein for the mutein is des ala 1 ser 125 IL-2.
Claim 4
Dependent Claim
22 words
A recombinant expression vector which comprises the DNA sequence of claim 2 operably linked to control sequences compatible with a suitable host.
Claim 5
Dependent Claim
22 words
A recombinant expression vector which comprises the DNA sequence of claim 1 operably linked to control sequences compatible with a suitable host.
Claim 6
Dependent Claim
12 words
The recombinant expression vector of claim 4 wherein the host is yeast.
Claim 7
Dependent Claim
25 words
The recombinant expression vector of claim 5 which comprises a gene coding for ala 104 IL-2 which is designated pPM43 and has ATCC No. 53,356.
Claim 8
Dependent Claim
16 words
The recombinant expression vector of clam 4 which is pSY3001 wherein the host is E. coli.
Claim 9
Dependent Claim
16 words
The recombinant expression vector of clam 4 which is pSY3001 wherein the host is E. coli.
Claim 10
Dependent Claim
10 words
Host cells transformed with the expression vector of claim 3.
Claim 11
Dependent Claim
10 words
Host cells transformed with the expression vector of claim 4.
Claim 12
Dependent Claim
27 words
The recombinant DNA sequence of claim 1 wherein the conservative amino acid sequence is selected from the group consisting of alanine, serine, leucine, isoleucine, glutamate and valine.
Claim 13
Dependent Claim
455 words
The recombinant DNA sequence of claim 1, which DNA sequence encodes for an oxidation-resistant mutein selected from the group consisting of; ala 104 ser 125 IL-2, ala 104 IL-2, ala 104 ala 125 IL-2, val 104 ser 125 IL-2, val 104 IL-2, val 104 ala 125 IL-2, des-ala 1 ala 104 ser 125 IL-2, des-ala 1 ala 104 IL-2, des-ala 1 ala 104 ala 125 IL-2, des-ala 1 val 104 ser 125 IL-2, des-ala 1 val 104 IL-2, des-ala 1 val 104 ala 125 IL-2, des-ala 1 des-pro 2 ala 104 ser 125 IL-2, des-ala 1 des-pro 2 ala 104 IL-2, des-ala 1 des-pro 2 ala 104 ala 125 IL-2, des-ala 1 des-pro 2 val 104 ser 125 IL-2, des-ala 1 des-pro 2 val 104 IL-2, des-ala 1 des-pro 2 val 104 ala 125 IL-2, des-ala 1 des-pro 2 des-thr 3 ala 104 ser 125 IL-2, des-ala 1 des-pro 2 des-thr 3 ala 104 IL-2, des-ala 1 des-pro.sub. 2 des-thr 3 ala 104 ala 125 IL-2, des-ala 1 des-pro 2 des-thr 3 val 104 ser 125 IL-2, des-ala 1 des-pro 2 des-thr 3 val 104 IL-2, des-ala 1 des-pro 2 des-thr 3 val 104 ala 125 IL-2, des-ala 1 des-pro 2 des-thr 3 des-ser 4 ala 104 ser 125 IL-2, des-ala 1 des-pro 2 des-thr 3 des-ser 4 ala 104 IL-2, des-ala 1 des-pro 2 des-thr 3 des-ser 4 ala 104 ala 125 IL-2, des-ala 1 des-pro 2 des-thr 3 des-ser 4 val 104 ser 125 IL-2, des-ala 1 des-pro 2 des-thr 3 des-ser 4 val 104 IL-2, des-ala 1 des-pro 2 des-thr 3 des-ser 4 val 104 ala 125 IL-2, des-ala 1 des-pro 2 des-thr 3 des-ser 4 des-ser 5 ala 104 ser 125 IL-2, des-ala 1 des-pro 2 des-thr 3 des-ser 4 des-ser 5 ala 104 IL-2, des-ala 1 des-pro 2 des-thr 3 des-ser 4 des-ser 5 ala 104 ala 125 IL-2, des-ala 1 des-pro 2 des-thr 3 des-ser 4 des-ser 5 val 104 ser 125 IL-2, des-ala 1 des-pro 2 des-thr 3 des-ser 4 des-ser 5 val 104 IL-2, des-ala 1 des-pro 2 des-thr 3 des-ser 4 des-ser 5 val 104 ala 125 IL-2, des-ala 1 des-pro 2 des-thr 3 des-ser 4 des-ser 5 des-ser 6 ala 104 ala 125 IL-2, des-ala 1 des-pro 2 des-thr 3 des-ser 4 des-ser 5 des-ser 6 ala 104 IL-2, des-ala 1 des-pro 2 des-thr 3 des-ser 4 des-ser 5 des-ser 6 ala 104 ser 125 IL-2, des-ala 1 des-pro 2 des-thr 3 des-ser 4 des-ser 5 des-ser 6 val 104 ser 125 IL-2, des-ala 1 des-pro 2 des-thr 3 des-ser 4 des-ser 5 des-ser 6 val 104 IL-2, des-ala 1 des-pro 2 des-thr 3 des-ser 4 des-ser 5 des-ser 6 val 104 ala 125 IL-2.
Claim 14
Dependent Claim
54 words
The recombinant DNA sequence of claim 1 which DNA sequence encodes for an oxidation-resistant mutein selected from the group consisting of; ala 104 ser 125 IL-2, ala 104 IL-2, ala 104 ala 125 IL-2, des-ala 1 ala 104 IL-2, des-ala 1 ala 104 ser 125 IL-2, or des-ala 1 ala 104 ala 125 IL-2.
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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) | 1991-06-21 | 1991-12-21 | 1992-06-21 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1995-06-21 | 1995-12-21 | 1996-06-21 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1999-06-21 | 1999-12-21 | 2000-06-21 | $7700.00 | Window Closed / Lapsed |