US4472509A
Expired / Lapsed
US
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Metal chelate conjugated monoclonal antibodies
Assignee: Johns Hopkins University
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Filing: 1982-06-07
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Granted: 1984-09-18
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Expires: 2002-06-07
Abstract
Abstract Methods of manufacturing and purifying metal chelate conjugated monoclonal antibodies are described. The chelated metal may be one which emits alpha, beta or gamma radiation, or positrons. Alternatively, the metal can be one which is fluorogenic or paramagnetic. The conjugates are suited for diagnostic and therapeutic uses.
Inventors
- 👤 Otto A. Gansow
- 👤 Mette Strand
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-24 14:24
CPC Classifications
A; A61; A61K; A61K51/00; A61K51/02; A61K51/04; A61K51/08; A61K51/10; A61K51/1093; C; C07; C07K; C07K16/00; C07K16/08; C07K16/10; C07K16/112; C07K16/18; C07K16/28; C07K16/30; C07K16/3015; A61K2121/00; Y; Y10; Y10S; Y10S436/00; Y10S436/804; Y10S436/811; Y10S436/813; Y10S436/819
Claims Analysis
Total Claims: 34
Claim 1
Independent Claim
67 words
A method of purifying metal chelate conjugated monoclonal antibodies comprising passing an aqueous solution containing metal chelate conjugated monoclonal antibodies through a chromatography column, said column having at least two different layers, an initial layer selected from a group consisting of an ion retardation resin, an anion exchange resin, a cation exchange resin and a chelating ion exchange resin, and a final layer comprising a sizing matrix.
Claim 2
Independent Claim
67 words
A method of purifying metal chelate conjugated monoclonal antibodies comprising passing an aqueous solution containing metal chelate conjugated monoclonal antibodies through a chromatography column, said column having at least two different layers, an initial layer selected from a group consisting of an ion retardation resin, an anion exchange resin, a cation exchange resin and a chelating ion exchange resin, and a final layer comprising a sizing matrix.
Claim 3
Dependent Claim
30 words
The method of claim 1 wherein an initial layer of an ion retardation resin, a second layer of a cation exchange resin and a final sizing matrix layer are employed.
Claim 4
Dependent Claim
16 words
The method of claim 1 wherein said chelate conjugated monoclonal antibodies are prepared from diethylenetriaminepentaacetic acid.
Claim 5
Independent Claim
55 words
A method of preparing metal chelate conjugated monoclonal antibodies comprising adding a metal salt to an aqueous solution of a chelate conjugated monoclonal antibodies and a buffer, said solution being maintained at a pH within the range of about 3.2 to about 9 throughout chelation without the addition of a strong acid or strong base.
Claim 6
Independent Claim
55 words
A method of preparing metal chelate conjugated monoclonal antibodies comprising adding a metal salt to an aqueous solution of a chelate conjugated monoclonal antibodies and a buffer, said solution being maintained at a pH within the range of about 3.2 to about 9 throughout chelation without the addition of a strong acid or strong base.
Claim 7
Dependent Claim
11 words
The method of claim 4 wherein said buffer is citric acid.
Claim 8
Dependent Claim
23 words
The method of claim 4 wherein said chelation is carried out at a pH within the range of about 3.2 to about 3.5.
Claim 9
Dependent Claim
16 words
The method of claim 4 wherein said chelate conjugated monoclonal antibodies are prepared from diethylenetriaminepentaacetic acid.
Claim 10
Independent Claim
194 words
A method for preparing metal chelate conjugated monoclonal antibodies comprising: (a) contacting a carboxycarbonic anhydride of diethylenetriaminepentaacetic acid with an aqueous solution of monoclonal antibodies at a pH maintained in the range from about 6 to about 7.2 to produce chelate conjugated monoclonal antibodies; (b) recovering said chelate conjugated monoclonal antibodies; (c) adding a metal salt to an aqueous solution of said recovered chelate conjugated monoclonal antibodies and a buffer to form metal chelate conjugated antibodies, said solution being maintained at a pH within the range of from about 3.2 to about 9 throughout chelation without the addition of a strong acid or strong base; (d) passing the aqueous solution containing said metal chelate conjugated monoclonal antibodies through a chromatography column, said column having one or more layers selected from the group consisting of an ion retardation resin, an anion exchange resin, a cation exchange resin and a chelating ion exchange resin, and a final layer comprising a sizing matrix; and (e) recovering from said column a solution of metal chelate conjugated antibodies having at least about 80% of said metal contained in said solution complexed by said chelate conjugated to said monoclonal antibodies.
Claim 11
Independent Claim
194 words
A method for preparing metal chelate conjugated monoclonal antibodies comprising: (a) contacting a carboxycarbonic anhydride of diethylenetriaminepentaacetic acid with an aqueous solution of monoclonal antibodies at a pH maintained in the range from about 6 to about 7.2 to produce chelate conjugated monoclonal antibodies; (b) recovering said chelate conjugated monoclonal antibodies; (c) adding a metal salt to an aqueous solution of said recovered chelate conjugated monoclonal antibodies and a buffer to form metal chelate conjugated antibodies, said solution being maintained at a pH within the range of from about 3.2 to about 9 throughout chelation without the addition of a strong acid or strong base; (d) passing the aqueous solution containing said metal chelate conjugated monoclonal antibodies through a chromatography column, said column having one or more layers selected from the group consisting of an ion retardation resin, an anion exchange resin, a cation exchange resin and a chelating ion exchange resin, and a final layer comprising a sizing matrix; and (e) recovering from said column a solution of metal chelate conjugated antibodies having at least about 80% of said metal contained in said solution complexed by said chelate conjugated to said monoclonal antibodies.
Claim 12
Dependent Claim
41 words
The method of claim 8 wherein said carboxycarbonic anhydride of diethylenetriaminepentaacetic acid is the reaction product of an amine diethylenetriaminepentaacetic acid salt with an ester of a haloformic acid in an organic solvent at a temperature less than about -20° C.
Claim 13
Dependent Claim
14 words
The method of claim 9 wherein said ester of a haloformic acid is isobutylchloroformate.
Claim 14
Dependent Claim
27 words
The method of claim 8 wherein said metal salt is added to said aqueous chelate conjugated monoclonal antibody solution in an aqueous buffered solution containing ascorbic acid.
Claim 15
Dependent Claim
40 words
An aqueous solution of metal chelate conjugated monoclonal antibodies wherein the metal ion is in at least the +3 valence state, and in which at least about 94% of said metal is complexed by the chelate portion of said conjugate.
Claim 16
Dependent Claim
40 words
An aqueous solution of metal chelate conjugated monoclonal antibodies wherein the metal ion is in at least the +3 valence state, and in which at least about 94% of said metal is complexed by the chelate portion of said conjugate.
Claim 17
Dependent Claim
22 words
The solution of claim 12 wherein at least about 97% of said metal is complexed by the chelate portion of said conjugate.
Claim 18
Dependent Claim
19 words
The solution of claim 12 wherein said conjugated antibodies retain at least about 80% of their activity and specificity.
Claim 19
Dependent Claim
19 words
The solution of claim 12 wherein said conjugated antibodies retain at least about 95% of their activity and specificity.
Claim 20
Dependent Claim
19 words
The solution of claim 12 wherein said conjugated antibodies retain at least about 90% of their activity and specificity.
Claim 21
Dependent Claim
17 words
The solution of claim 12 wherein said conjugated antibodies retain substantially all of their activity and specificity.
Claim 22
Dependent Claim
14 words
The solution of claim 12 wherein said conjugated antibodies are formed employing diethylenetriaminepentaacetic acid.
Claim 23
Dependent Claim
17 words
The solution of claim 18 wherein said conjugated antibodies retain substantially all of their activity and specificity.
Claim 24
Independent Claim
123 words
A method for preparing chelate conjugated monoclonal antibodies comprising: reacting an ester of a haloformic acid with an amine salt of diethylenetriaminepentaacetic acid in an organic solvent to produce carboxycarbonic anhydride of diethylenetriaminepentaacetic acid, said reaction carried out at a temperature sufficiently low to precipitate substantially all of the haloamine salt by-product; recovering said carboxycarbonic anhydride of diethylenetriaminepentaacetic acid by separating said organic solvent containing the anhydride in solution from said precipitated haloamine salt by-product; contacting said carboxycarbonic anhydride of diethylenetriaminepentaacetic acid with an aqueous solution containing monoclonal antibodies at a pH maintained in a range from about 6.0 to about 7.2 to produce chelate conjugated monoclonal antibodies; and recovering said chelate conjugated monoclonal antibodies having their biological activity and specificities substantially retained.
Claim 25
Independent Claim
123 words
A method for preparing chelate conjugated monoclonal antibodies comprising: reacting an ester of a haloformic acid with an amine salt of diethylenetriaminepentaacetic acid in an organic solvent to produce carboxycarbonic anhydride of diethylenetriaminepentaacetic acid, said reaction carried out at a temperature sufficiently low to precipitate substantially all of the haloamine salt by-product; recovering said carboxycarbonic anhydride of diethylenetriaminepentaacetic acid by separating said organic solvent containing the anhydride in solution from said precipitated haloamine salt by-product; contacting said carboxycarbonic anhydride of diethylenetriaminepentaacetic acid with an aqueous solution containing monoclonal antibodies at a pH maintained in a range from about 6.0 to about 7.2 to produce chelate conjugated monoclonal antibodies; and recovering said chelate conjugated monoclonal antibodies having their biological activity and specificities substantially retained.
Claim 26
Dependent Claim
29 words
The method of claim 12 wherein the molar ratio of said carboxycarbonic anhydride of diethylenetriaminepentaacetic acid to said monoclonal antibodies in said contacting step is at least about 300:1.
Claim 27
Dependent Claim
25 words
The method of claim 20 wherein the concentration of said carboxycarbonic anhydride of diethylenetriaminepentaacetic acid recovered in the organic solvent is at least about 0.25M.
Claim 28
Dependent Claim
14 words
The method of claim 20 wherein said ester of a haloformic acid is isobutylchloroformate.
Claim 29
Dependent Claim
22 words
The method of claim 20 wherein said reaction temperature is in the range of from about -20° C. to about -70° C.
Claim 30
Dependent Claim
18 words
The solution of claim 13 wherein said conjugated antibodies retain at least 80% of their activity and specificity.
Claim 31
Dependent Claim
18 words
The solution of claim 13 wherein said conjugated antibodies retain at least 90% of their activity and specificity.
Claim 32
Dependent Claim
18 words
The solution of claim 13 wherein said conjugated antibodies retain at least 95% of their activity and specificity.
Claim 33
Dependent Claim
17 words
The solution of claim 13 wherein said conjugated antibodies retain substantially all of their activity and specificity.
Claim 34
Dependent Claim
14 words
The solution of claim 13 wherein said conjugated antibodies are formed employing diethylenetriaminepentaacetic acid.
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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) | 1987-09-18 | 1988-03-19 | 1988-09-18 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1991-09-18 | 1992-03-19 | 1992-09-18 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1995-09-18 | 1996-03-19 | 1996-09-18 | $7700.00 | Window Closed / Lapsed |