US4780210A
Expired / Lapsed
US
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Tangential flow affinity ultrafiltration
Assignee: Canada, AS REPRESENTED BY MINISTER OF NATIONAL DEFENCE
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Filing: 1987-01-07
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Granted: 1988-10-25
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Expires: 2007-01-07
Abstract
Abstract Biological macromolecules are separated from liquid mixtures in which they are contained, in substantially pure form, by a process involving tangential flow ultrafiltration of the liquid mixture in the presence of an affinity gel which binds selectively to the biological macromolecule to be recovered in pure form, in either the positive affinity absorption mode or the negative affinity absorption mode. Thus a mixture containing the biological macromolecule of interest, such as hemoglobin or (oxy)hemoglobin is first mixed with an affinity gel, such as agarose-ATP, which selectively binds to the hemoglobin or (oxy)hemoglobin and then the liquid is subjected to tangential flow ultrafiltration, so that all components of the mixture except the gel-bond hemoglobin pass through the filter. In a second stage, the gel-bond hemoglobin is treated with a salt solution to displace the hemoglobin from the gel, and then the mixture is passed again over a tangential flow ultrafiltration membrance, so that the pure hemoglobin is separated from the gel, and collected in filtrate. The affinity gel which remains in retentate can then be regenerated and re-used.
Inventors
- 👤 Jen-Chang Hsia
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-10 01:57
CPC Classifications
C; C07; C07K; C07K1/00; C07K1/14; C07K1/34; C07K14/00; C07K14/795; C07K14/805
Claims Analysis
Total Claims: 16
Claim 1
Independent Claim
90 words
A process of separating a first biological macromolecule from a second biological macromolecule which comprises treating a mixture containing both of said first and second biological macromolecules with a macromolecular affinity substance having ligands which endow biological binding specificity to one of said first and second macromolecules but not to the other thereof, and subsequently subjecting the mixture containing the affinity substance-bound macromolecules to tangential flow ultrafiltration, so as to obtain affinity substance-bound macromolecules in the retentate of said ultrafiltration and non-affinity substance-bound macromolecules in the filtrate of said ultrafiltration.
Claim 2
Dependent Claim
90 words
A process of separating a first biological macromolecule from a second biological macromolecule which comprises treating a mixture containing both of said first and second biological macromolecules with a macromolecular affinity substance having ligands which endow biological binding specificity to one of said first and second macromolecules but not to the other thereof, and subsequently subjecting the mixture containing the affinity substance-bound macromolecules to tangential flow ultrafiltration, so as to obtain affinity substance-bound macromolecules in the retentate of said ultrafiltration and non-affinity substance-bound macromolecules in the filtrate of said ultrafiltration.
Claim 3
Dependent Claim
23 words
The process of claim 1 wherein one of said first biological macromolecule and said second biological macromolecule is hemoglobin or a derivative thereof.
Claim 4
Dependent Claim
32 words
The process of claim 2 wherein the affinity gel is an agarose gel having a selective affinity binding anion attached thereto through a space or chemical group of at least 5 angstroms.
Claim 5
Dependent Claim
36 words
The process of claim 3 wherein the selective affinity binding anion is an organic phosphate, a nucleic acid phosphate, an inositol phosphate, a carbohydrate phosphate, a carbohydrate sulfate, a carbohydrate monocarboxylate, a carbohydrate polycarboxylate or diphosphoglycerate.
Claim 6
Dependent Claim
52 words
The process of claim 1 wherein, subsequent to the tangential flow ultrafiltration, the affinity-substance-bound macromolecules in the retentate are chemically displaced from binding with the macromolecular affinity substance, and the mixture of macromolecular affinity substance and macromolecules so formed is subjected to tangential flow ultrafiltration to separate the macromolecular affinity substance therefrom.
Claim 7
Dependent Claim
31 words
The process of claim 5 wherein the macromolecular affinity substance is subsequently recycled to bind selectively to one of said first and second macromolecules from another batch of mixture containing them.
Claim 8
Dependent Claim
14 words
The process of 1 wherein the macromolecular affinity substance is an affinity chromatographic gel.
Claim 9
Dependent Claim
14 words
The process of 1 wherein the macromolecular affinity substance is an affinity chromatographic gel.
Claim 10
Dependent Claim
17 words
The process of claim 7 wherein the affinity gel is an agarose gel or a silica gel.
Claim 11
Dependent Claim
17 words
The process of claim 8 wherein said first and second biological macromolecules are hemoglobin and hemoglobin derivatives.
Claim 12
Dependent Claim
18 words
The process of claim 8 wherein the first and second biological macromolecules are a glycoprotein and a non-glycoprotein.
Claim 13
Dependent Claim
14 words
The process of claim 10 wherein the affinity gel is agarose bound to concanavalin-A.
Claim 14
Dependent Claim
16 words
The process of claim 1 wherein the macromolecular affinity substance is a soluble ligand-binding macromolecular substance.
Claim 15
Dependent Claim
17 words
The process of claim 1 wherein said first and second biological macromolecules are hemoglobin and hemoglobin derivatives.
Claim 16
Dependent Claim
19 words
The process of claim 1 wherein one of said first biological macromolecule and said second biological macromolecule is oxyhemoglobin.
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) | 1991-10-25 | 1992-04-25 | 1992-10-25 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1995-10-25 | 1996-04-25 | 1996-10-25 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1999-10-25 | 2000-04-25 | 2000-10-25 | $7700.00 | Window Closed / Lapsed |