US4801687A
Expired / Lapsed
US
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Monoclonal antibody purification process using protein A
Assignee: BIOPROBE INTERNATIONAL Inc 2842 WALNUT AVENUE SUITE C A DE CORP
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Filing: 1987-12-30
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Granted: 1989-01-31
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Expires: 2007-12-30
Abstract
Abstract Immunoglobins are purified by adsorption upon an immobilized protein A adsorbent using a buffer having a pH of 7.5 to 10 and containing a combination of monovalent cations and polybasic anions in a concentration of about 0.6M to 1.75M.
Inventors
- 👤 That T. Ngo
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-08 15:19
CPC Classifications
C; C07; C07K; C07K16/00; C07K16/06; C07K16/065; Y; Y10; Y10S; Y10S530/00; Y10S530/808; Y10S530/809; Y10S530/827; Y10S530/829; Y10S530/83
Claims Analysis
Total Claims: 21
Claim 1
Independent Claim
143 words
A process for the purification of immunoglobulins which comprises: mixing a medium containing immunoglobulins with a buffer solution having a pH in the range of about pH 7.5 to pH 10 and containing a combination of monovalent cations and polybasic anions in a concentration of about 0.6M to 1.75M to provide a buffered immunoglobulin medium; contacting said buffered immunoglobulin medium with an immobilized protein A adsorbent to adsorb the immunoglobulins present in said buffered immunoglobulin medium upon said immobilized protein A adsorbent; washing the immobilized protein A adsorbent having immunoglobulins adsorbed thereon with said buffer solution; contacting said immobilized protein A adsorbent having immunoglobulins adsorbed thereon with a buffer solution having a pH in the range of about pH 3 to pH 6 to remove the adsorbed immunoglobulins from the immobilized protein A adsorbent; and recovering the removed immunoglobulins in substantially pure form.
Claim 2
Dependent Claim
143 words
A process for the purification of immunoglobulins which comprises: mixing a medium containing immunoglobulins with a buffer solution having a pH in the range of about pH 7.5 to pH 10 and containing a combination of monovalent cations and polybasic anions in a concentration of about 0.6M to 1.75M to provide a buffered immunoglobulin medium; contacting said buffered immunoglobulin medium with an immobilized protein A adsorbent to adsorb the immunoglobulins present in said buffered immunoglobulin medium upon said immobilized protein A adsorbent; washing the immobilized protein A adsorbent having immunoglobulins adsorbed thereon with said buffer solution; contacting said immobilized protein A adsorbent having immunoglobulins adsorbed thereon with a buffer solution having a pH in the range of about pH 3 to pH 6 to remove the adsorbed immunoglobulins from the immobilized protein A adsorbent; and recovering the removed immunoglobulins in substantially pure form.
Claim 3
Dependent Claim
31 words
A process according to claim 1 wherein the contacting of said buffered immunoglobulin medium with said immobilized protein A adsorbent is accomplished in a column of said immobilized protein A adsorbent.
Claim 4
Dependent Claim
16 words
A process according to claim 1 wherein said medium containing immunoglobulins is a normal mammalian serum.
Claim 5
Dependent Claim
16 words
A process according to claim 1 wherein said medium containing immunoglobulins is an immune mammalian serum.
Claim 6
Dependent Claim
13 words
A process according to claim 1 wherein said medium is a mammalian plasma.
Claim 7
Dependent Claim
13 words
A process according to claim 1 wherein said medium is mammalian ascites fluid.
Claim 8
Dependent Claim
14 words
A process according to claim 1 wherein said medium is obtained from a hybridoma.
Claim 9
Dependent Claim
14 words
A process according to claim 1 wherein said medium is a tissue culture fluid.
Claim 10
Dependent Claim
27 words
A process according to claim 1 wherein said buffer solution having a pH in the range of about pH 7.5 to pH 10 is a glycine buffer.
Claim 11
Dependent Claim
27 words
A process according to claim 1 wherein said buffer solution having a pH in the range of about pH 7.5 to pH 10 is a borate buffer.
Claim 12
Dependent Claim
29 words
A process according to claim 1 wherein said buffer solution having a pH in the range of about pH 7.5 to pH 10 is a tris (hydroxymethyl) aninomethane buffer.
Claim 13
Dependent Claim
34 words
A process according to claim 1 wherein said buffer solution having a pH in the range of about pH 7.5 to pH 10 has a concentration in the range of about 0.01M to 0.25M.
Claim 14
Dependent Claim
20 words
A process according to claim 1 wherein said immobilized protein A adsorbent is protein A chemically bonded to cross-linked agarose.
Claim 15
Dependent Claim
26 words
A process according to claim 1 wherein said immobilized protein A adsorbent is protein A chemically bonded to a 2-fluoro-1-methylpyridinium toluene-4-sulfonate activated hydroxyl group-containing polymeric carrier.
Claim 16
Dependent Claim
28 words
A process according to claim 1 wherein said buffer solution having a pH in the range of about pH 3 to pH 6 is an acetic acid-acetate buffer.
Claim 17
Dependent Claim
34 words
A process according to claim 1 wherein said buffer solution having a pH in the range of about pH 3 to pH 6 has a concentration in the range of about 0.01M to 0.25M.
Claim 18
Dependent Claim
24 words
A process according to claim 1 wherein said buffer solution contains potassium ions and phosphate ions in a concentration of about 1.0M to 1.5M.
Claim 19
Dependent Claim
24 words
A process according to claim 1 wherein said buffer solution contains ammonium ions and phosphate ions in a concentration of about 1.0M to 1.5M.
Claim 20
Dependent Claim
24 words
A process according to claim 1 wherein said buffer solution contains ammonium ions and sulfate ions in a concentration of about 1.0M to 1.5M.
Claim 21
Dependent Claim
24 words
A process according to claim 1 wherein said buffer solution contains sodium ions and sulfate ions in a concentration of about 1.0M to 1.25M.
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) | 1992-01-31 | 1992-08-01 | 1993-01-31 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1996-01-31 | 1996-08-01 | 1997-01-31 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2000-01-31 | 2000-08-01 | 2001-01-31 | $7700.00 | Window Closed / Lapsed |