US4889916A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Protein label and drug delivery system
Assignee: Johns Hopkins University
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Filing: 1988-02-01
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Granted: 1989-12-26
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Expires: 2008-02-01
Abstract
Abstract A bridging molecule carrying a drug, or a label such as a fluorophore, which adds across disulfide bonds of molecules, particularly proteins, and methods of manfacturing and using the bridging molecules, are disclosed. The bridging molecule is reactive with sulfhydryl groups formed by the reduction of disulfide bonds of the protein. The functional groups of the bridging molecules are typically -SH groups.
Inventors
- 👤 Beverly Packard
- 👤 Michael Edidin
- 👤 Akira Komoriya
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-01 15:37
CPC Classifications
C; C07; C07K; C07K1/00; C07K1/107; C07K1/1072; C07K1/1077; A; A61; A61K; A61K47/00; A61K47/50; A61K47/51; A61K47/68; A61K47/6889; C07D; C07D311/00; C07D311/02; C07D311/78; C07D311/80; C07D311/82; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/531; G01N33/532; G01N33/533
Claims Analysis
Total Claims: 13
Claim 1
Independent Claim
91 words
A method of conjugating a protein with a bridging compound having the formula ##STR8## wherein X and Y comprise separate functional groups reactive with sulfhydryl groups formed by reduction of disulfide bonds of a protein molecule and A comprises a drug, dye or label portion which is unreactive with sulfhydryl groups, said method comprising the steps of: (a) reducing the disulfide bonds of said protein; (b) reacting the bridging compound across the reduced disulfide bonds of said protein; and (c) oxidizing or alkylating the reduced disulfide bonds remaining after step (b).
Claim 2
Independent Claim
91 words
A method of conjugating a protein with a bridging compound having the formula ##STR8## wherein X and Y comprise separate functional groups reactive with sulfhydryl groups formed by reduction of disulfide bonds of a protein molecule and A comprises a drug, dye or label portion which is unreactive with sulfhydryl groups, said method comprising the steps of: (a) reducing the disulfide bonds of said protein; (b) reacting the bridging compound across the reduced disulfide bonds of said protein; and (c) oxidizing or alkylating the reduced disulfide bonds remaining after step (b).
Claim 3
Dependent Claim
24 words
A method as in claim 1 wherein all the steps are carried out at between room temperature and the denaturation temperature of said protein.
Claim 4
Dependent Claim
26 words
A method as in claim 1 wherein the reacting step includes the step of adding about a 100-fold excess of the bridging compound to said protein.
Claim 5
Dependent Claim
23 words
A method as in claim 3 wherein the reacting step includes the steps of incubating the bridging compound-protein mixture for about 10 minutes.
Claim 6
Dependent Claim
27 words
A method as in claim 1 wherein the oxidizing step includes the steps of bubbling oxygen gas through the solution, and dropping the pH to about 5.5.
Claim 7
Dependent Claim
20 words
A method as in claim 1 wherein the reducing step includes the step of reducing the disulfide bonds with dithiothreitol.
Claim 8
Dependent Claim
22 words
A method as in claim 1 wherein the reducing step includes the step of reducing the disulfide bonds with 2 mercapto ethanol.
Claim 9
Dependent Claim
27 words
A method as in claim 1 wherein the reducing step includes the step of mixing dithiothreitol with said protein at a ratio of from 1:1 to 10:1.
Claim 10
Dependent Claim
20 words
A method as in claim 1 wherein the reducing step includes the step of adding an excess of ethylenediaminetetraacetic acid.
Claim 11
Dependent Claim
26 words
A method as in claim 1 wherein the reducing step includes the step of adjusting the pH to about 8-10 and incubating for about 30 minutes.
Claim 12
Dependent Claim
25 words
A method as in claim 10 wherein the oxidizing step includes the step of keeping the solution at about room temperature for about 22 hours.
Claim 13
Dependent Claim
14 words
A method according to claim 1 where X and Y are halogen or sulfhydryl.
References Cited (Backward Citations)
US4618492A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1992-12-25 | 1993-06-26 | 1993-12-26 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1996-12-25 | 1997-06-26 | 1997-12-26 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2000-12-25 | 2001-06-26 | 2001-12-26 | $7700.00 | Window Closed / Lapsed |