US4485017A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Isolation of human interferon by immunosorbent and high performance liquid chromatography
Assignee: Novartis Vaccines and Diagnostics Inc
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Filing: 1982-12-22
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Granted: 1984-11-27
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Expires: 2002-12-22
Abstract
Abstract An improved process for the isolation and purification of HIFNs is disclosed wherein a partially purified preparation of the HIFN is sequentially passed through an antibody affinity column and a reversed-phase high performance liquid chromatographic column. Organic solvents used during the elution are extracted and the protein concentrated for subsequent use.
Inventors
- 👤 Yin H. Tan
- 👤 Heidi Smith-Johannsen
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-04 15:16
CPC Classifications
B; B01; B01D; B01D15/00; B01D15/08; B01D15/26; B01D15/38; B01D15/3804; B01D15/3809; C; C07; C07K; C07K14/00; C07K14/435; C07K14/52; C07K14/555; C07K14/565; Y; Y10; Y10S; Y10S435/00; Y10S435/811
Claims Analysis
Total Claims: 15
Claim 1
Independent Claim
70 words
A process for the isolation and purification of a lipophilic interferon protein comprising (a) applying a solution of said protein to an immunosorbent adsorption affinity column; (b) eluting said protein from said affinity column with an equilibrating buffer; (c) passing the eluate from said affinity column containing said protein through a reversed-phase high performance liquid chromatographic column; and (d) eluting said protein from said chromatographic column with an organic solvent.
Claim 2
Dependent Claim
70 words
A process for the isolation and purification of a lipophilic interferon protein comprising (a) applying a solution of said protein to an immunosorbent adsorption affinity column; (b) eluting said protein from said affinity column with an equilibrating buffer; (c) passing the eluate from said affinity column containing said protein through a reversed-phase high performance liquid chromatographic column; and (d) eluting said protein from said chromatographic column with an organic solvent.
Claim 3
Dependent Claim
27 words
A process according to claim 1, wherein said equilibrating buffer is a plurality of buffers and said eluting is carried out serially with said plurality of buffers.
Claim 4
Dependent Claim
23 words
A process according to claim 1, wherein the pH of said equilibrating buffer is in the range of about 2.0 to about 3.0.
Claim 5
Dependent Claim
30 words
A process according to claim 1, wherein the organic solvent of step (d) is selected from the group consisting of methanol, ethanol, n-propanol, 2-propanol, dimethyl formamide, acetonitrile, acetone and dioxane.
Claim 6
Dependent Claim
12 words
A process according to claim 1, wherein said interferon is human interferon.
Claim 7
Dependent Claim
14 words
A process according to claim 5, wherein said human interferon is human fibroblast interferon.
Claim 8
Dependent Claim
11 words
A process according to claim 4, wherein said solvent is n-propanol.
Claim 9
Dependent Claim
17 words
A process according to claim 7, wherein said elution with n-propanol is used over a linear gradient.
Claim 10
Dependent Claim
21 words
A process according to claim 7 further comprising extracting said n-propanol with cyclohexane and removing said cyclohexane with a stream nitrogen.
Claim 11
Dependent Claim
94 words
A process for the isolation and purification of biologically active human interferon comprising: (a) applying a solution of ammonium sulfate precipitated HIFN to an antibody affinity column; (b) eluting said HIFN from said affinity column with a buffer; (c) ultrafiltering the eluate; (d) injecting said ultrafiltered HIFN solution onto a reverse-phase high performance liquid chromatographic column; (e) eluting said HIFN from said chromatographic column with a linear gradient flow of n-propanol; (f) extracting said n-propanol from the eluate with cyclohexane; and (g) remvoing residual cyclohexane from said HIFN solution with a stream of nitrogen.
Claim 12
Dependent Claim
94 words
A process for the isolation and purification of biologically active human interferon comprising: (a) applying a solution of ammonium sulfate precipitated HIFN to an antibody affinity column; (b) eluting said HIFN from said affinity column with a buffer; (c) ultrafiltering the eluate; (d) injecting said ultrafiltered HIFN solution onto a reverse-phase high performance liquid chromatographic column; (e) eluting said HIFN from said chromatographic column with a linear gradient flow of n-propanol; (f) extracting said n-propanol from the eluate with cyclohexane; and (g) remvoing residual cyclohexane from said HIFN solution with a stream of nitrogen.
Claim 13
Dependent Claim
87 words
A process for preparing human fibroblast interferon which process comprises (a) applying a solution at neutral pH of the human fibroblast interferon to an immunosorbent affinity column comprising antibodies to human fibroblast interferon immobilized on a solid support; (b) eluting the human fibroblast interferon with a buffer of pH about 2-3; (c) applying the human fibroblast interferon containing fractions from (b) to a reverse phase, high performance liquid chromatography column; (d) eluting the human fibroblast interferon from said chromatography column with a linear gradient flow of n-propanol.
Claim 14
Dependent Claim
87 words
A process for preparing human fibroblast interferon which process comprises (a) applying a solution at neutral pH of the human fibroblast interferon to an immunosorbent affinity column comprising antibodies to human fibroblast interferon immobilized on a solid support; (b) eluting the human fibroblast interferon with a buffer of pH about 2-3; (c) applying the human fibroblast interferon containing fractions from (b) to a reverse phase, high performance liquid chromatography column; (d) eluting the human fibroblast interferon from said chromatography column with a linear gradient flow of n-propanol.
Claim 15
Dependent Claim
25 words
The process of claim 11, which further includes extracting the n-propanol from the eluate with cyclohexane and removing residual cyclohexane with a stream of nitrogen.
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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) | 1987-11-27 | 1988-05-28 | 1988-11-27 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1991-11-27 | 1992-05-28 | 1992-11-27 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1995-11-27 | 1996-05-28 | 1996-11-27 | $7700.00 | Window Closed / Lapsed |