US4533496A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Method of isolating monoclonal antibodies from hybridoma cultures
Assignee: Monsanto Co
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Filing: 1984-05-08
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Granted: 1985-08-06
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Expires: 2004-05-08
Abstract
Abstract Monoclonal antibodies are isolated from spent culture medium of the in vitro growth of hybridoma cells by treating said spent cell culture medium or a concentrate thereof with a water-insoluble, cross-linked polyelectrolyte copolymer.
Inventors
- 👤 Charles Lewis, Jr.
- 👤 Jitka V. Olander
- 👤 William R. Tolbert
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-03 12:25
CPC Classifications
C; C07; C07K; C07K16/00; Y; Y10; Y10S; Y10S530/00; Y10S530/808; Y10S530/809
Claims Analysis
Total Claims: 18
Claim 1
Independent Claim
68 words
A method of isolating monoclonal antibodies from spent culture medium of the in vitro growth of hybridoma cells comprising treating said spent cell culture medium or a concentrate thereof by adsorption with a water-insoluble, cross-linked polyelectrolyte copolymer of olefinically unsaturated monomer having from 2 to about 4 carbon atoms and α,β-unsaturated polycarboxylic acid or anhydride having from 4 to about 6 carbon atoms containing pendant diloweralkylaminoloweralkylimide functional groups.
Claim 2
Independent Claim
68 words
A method of isolating monoclonal antibodies from spent culture medium of the in vitro growth of hybridoma cells comprising treating said spent cell culture medium or a concentrate thereof by adsorption with a water-insoluble, cross-linked polyelectrolyte copolymer of olefinically unsaturated monomer having from 2 to about 4 carbon atoms and α,β-unsaturated polycarboxylic acid or anhydride having from 4 to about 6 carbon atoms containing pendant diloweralkylaminoloweralkylimide functional groups.
Claim 3
Dependent Claim
26 words
The method of claim 1 in which the olefinically unaturated monomer is ethylene and the α,β- unsaturated polycarboxylic acid or anhydride is maleic acid or anhydride.
Claim 4
Dependent Claim
11 words
The method of claim 2 in which the diloweralkylaminoloweralkylimide is dimethylaminopropylimide.
Claim 5
Dependent Claim
24 words
The method of claim 3 in which the polyelectrolyte copolymer contains from about 3 mole % to about 100 mole % of said dimethylaminopropylimide.
Claim 6
Dependent Claim
18 words
The method of claim 1 including the additional step of desorption of protein adsorbed to said polyelectrolyte copolymer.
Claim 7
Dependent Claim
35 words
The method of claim 1 in which the adsorption is carried out at a pH of from about 6.5 to about 7.5 and an ionic salt concentration of from about 0.01 to about 0.15 molar.
Claim 8
Dependent Claim
35 words
The method of claim 5 in which the desorption is carried out at a pH of from about 4.7 to about 5.5 and an ionic salt concentration of from about 0.01 to about 0.15 molar.
Claim 9
Dependent Claim
35 words
The method of claim 6 in which the desorption is carried out at a pH of from about 4.7 to about 5.5 and an ionic salt concentration of from about 0.01 to about 0.15 molar.
Claim 10
Independent Claim
128 words
A method of isolating monoclonal antibodies from spent culture medium of the in vitro growth of hybridoma cells comprising treating said spent cell culture medium or a concentrate thereof by adsorption with a water-insoluble, cross-linked polyelectrolyte copolymer of ethylene and maleic acid or anhydride and containing from about 3 mole % to about 100 mole % of pendant dimethylaminopropylimide functional groups followed by desorption of protein adsorbed to said polyelectrolyte copolymer, said adsorption being carried out at a pH of from about 6.5 to about 7.5 and an ionic salt concentration of from about 0.01 to about 0.04 molar and said desorption being carried out at a pH of from about 4.7 to about 5.5 and a ionic salt concentration of from about 0.01 to about 0.04 molar.
Claim 11
Independent Claim
128 words
A method of isolating monoclonal antibodies from spent culture medium of the in vitro growth of hybridoma cells comprising treating said spent cell culture medium or a concentrate thereof by adsorption with a water-insoluble, cross-linked polyelectrolyte copolymer of ethylene and maleic acid or anhydride and containing from about 3 mole % to about 100 mole % of pendant dimethylaminopropylimide functional groups followed by desorption of protein adsorbed to said polyelectrolyte copolymer, said adsorption being carried out at a pH of from about 6.5 to about 7.5 and an ionic salt concentration of from about 0.01 to about 0.04 molar and said desorption being carried out at a pH of from about 4.7 to about 5.5 and a ionic salt concentration of from about 0.01 to about 0.04 molar.
Claim 12
Dependent Claim
17 words
The method of claim 5 including the additional step of subjecting the desorbed protein to salt precipitation.
Claim 13
Dependent Claim
17 words
The method of claim 8 including the additional step of subjecting the desorbed protein to salt precipitation.
Claim 14
Dependent Claim
13 words
The method of claim 10 in which the precipitant salt is ammonium sulfate.
Claim 15
Dependent Claim
13 words
The method of claim 11 in which the precipitant salt is ammonium sulfate.
Claim 16
Independent Claim
152 words
A method of isolating monoclonal antibodies from spent culture medium of the in vitro growth of hybridoma cells comprising treating said spent cell culture medium or a concentrate thereof by adsorption with a water-insoluble, cross-linked polyelectrolyte copolymer of ethylene and maleic acid or anhydride and containing from about 3 mole % to about 100 mole % of pendant dimethylaminopropylimide functional groups followed by desorption of protein adsorbed to said polyelectrolyte copolymer and thereafter subjecting said desorbed protein to salt precipitation, said adsorption being carried out at a pH of from about 6.5 to about 7.5 and an ionic salt concentration of from about 0.01 to about 0.04 molar, said desorption being carried out at a pH of from about 4.7 to about 5.5 and an ionic salt concentration of from about 0.01 to about 0.04 molar and in which the precipitant salt is from about 2 to about 2.4 molar ammonium sulfate.
Claim 17
Independent Claim
152 words
A method of isolating monoclonal antibodies from spent culture medium of the in vitro growth of hybridoma cells comprising treating said spent cell culture medium or a concentrate thereof by adsorption with a water-insoluble, cross-linked polyelectrolyte copolymer of ethylene and maleic acid or anhydride and containing from about 3 mole % to about 100 mole % of pendant dimethylaminopropylimide functional groups followed by desorption of protein adsorbed to said polyelectrolyte copolymer and thereafter subjecting said desorbed protein to salt precipitation, said adsorption being carried out at a pH of from about 6.5 to about 7.5 and an ionic salt concentration of from about 0.01 to about 0.04 molar, said desorption being carried out at a pH of from about 4.7 to about 5.5 and an ionic salt concentration of from about 0.01 to about 0.04 molar and in which the precipitant salt is from about 2 to about 2.4 molar ammonium sulfate.
Claim 18
Dependent Claim
11 words
Monoclonal antibody bound to polyelectrolyte copolymer as defined in claim 1.
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) | 1988-08-05 | 1989-02-04 | 1989-08-06 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1992-08-05 | 1993-02-04 | 1993-08-06 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1996-08-05 | 1997-02-04 | 1997-08-06 | $7700.00 | Window Closed / Lapsed |