US4411992A
Expired / Lapsed
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Process for preparing murine interleukin 2
Assignee: Immunex Corp
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Filing: 1981-04-14
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Granted: 1983-10-25
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Expires: 2001-04-14
Abstract
Abstract A process for preparing murine IL-2 from malignant neoplastic cells includes culturing murine leukemia or lymphoma cells in vitro in a protein containing medium supplemented with various additives. An optimum concentration of a T cell mitogen is added to the culture medium to stimulate maximum production of a supernate which contains IL-2. After a period of time, the supernate is collected and purified into more concentrated form. Phorbol myristate acetate may be added to a suboptimum concentration of the T cell mitogen to reduce the quantity of the mitogen required to produce maximum quantities of murine IL-2.
Inventors
- 👤 Steven Gillis
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-05 05:22
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/52; C07K14/54; C07K14/55; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/948
Claims Analysis
Total Claims: 27
Claim 1
Independent Claim
34 words
A process for the production of IL-2 from a murine malignant T-cell line, comprising culturing malignant murine T-cells in a culture medium containing a T-cell mitogen, and recovering the IL-2 from the culture medium.
Claim 2
Dependent Claim
34 words
A process for the production of IL-2 from a murine malignant T-cell line, comprising culturing malignant murine T-cells in a culture medium containing a T-cell mitogen, and recovering the IL-2 from the culture medium.
Claim 3
Dependent Claim
18 words
The process of claim 1, wherein said malignant murine T-cells are T leukemic cells or T lymphoma cells.
Claim 4
Dependent Claim
18 words
The process of claim 2, wherein said T lymphoma cells are from the LBRM-33 lymphoma murine T-cell line.
Claim 5
Dependent Claim
31 words
The process of claim 2 or 3, wherein the initial T leukemic or T lymphoma cell concentration is in the range of about 6×10 5 to 3×10 6 cells per milliliter.
Claim 6
Dependent Claim
21 words
The process of claim 2, wherein said malignant murine cells are clones of said T leukemic cells or T lymphoma cells.
Claim 7
Dependent Claim
20 words
The process of claim 5, wherein said clones of said T lymphoma cells are clones of LBRM-33 lymphoma murine T-cells.
Claim 8
Dependent Claim
33 words
The process of claim 5, or 6, wherein the initial T leukemic clone or T lymphoma clone cell concentration is in the range of about 6×10 5 to 3×10 6 cells per milliliter.
Claim 9
Dependent Claim
33 words
The process of claims 1, 2, 3, 5, or 6, wherein the T-cell mitogen in the culture medium is a compound selected from the group consisting of phytohemagglutinin, concanavalin A and pokeweed mitogen.
Claim 10
Dependent Claim
18 words
The process of claim 8, wherein the concentration of phytohemagglutinin mitogen is between 0.5 and 2.0% by volume.
Claim 11
Dependent Claim
20 words
The process of claim 8, wherein the concentration of concanavalin A mitogen is between 10 and 50 micrograms per milliliter.
Claim 12
Dependent Claim
15 words
The process of claim 8, wherein the culture medium further includes a phorbol ester derivative.
Claim 13
Dependent Claim
14 words
The process of claim 11, wherein the phorbol ester derivative includes phorbol myristate acetate.
Claim 14
Dependent Claim
22 words
The process of claim 12, wherein the phorbol myristate acetate is in a concentration of approximately 10 to 50 nanograms per milliliter.
Claim 15
Dependent Claim
24 words
The process of claim 8, wherein the culture medium further includes either Roswell Park Memorial Institute medium, Click's medium, or Dulbecco Modified Eagle Medium.
Claim 16
Dependent Claim
21 words
The process of claim 8, comprising recovering the IL-2 from the culture medium after approximately twenty-four to forty-eight hours of culturing.
Claim 17
Dependent Claim
36 words
The process of claim 1, 2, 3, 5, or 6, wherein the culture medium further contains a compound selected from a group consisting of Roswell Park Memorial Institute medium, Click's medium, or Dulbecco Modified Eagle Medium.
Claim 18
Dependent Claim
34 words
The process of claim 16, wherein the culture medium further contains one or more compounds selected from the group consisting of penicillin, streptomycin, glutamine, Hepes buffer, NaHCO 3 , fetal calf serum and 2-mercaptoethanol.
Claim 19
Dependent Claim
38 words
A process for producing IL-2 from a murine malignant T-cell line, comprising cloning malignant murine T-cells, culturing the cloned malignant murine T-cells in a culture medium containing a T-cell mitogen, and recovering the IL-2 from the culture medium.
Claim 20
Dependent Claim
38 words
A process for producing IL-2 from a murine malignant T-cell line, comprising cloning malignant murine T-cells, culturing the cloned malignant murine T-cells in a culture medium containing a T-cell mitogen, and recovering the IL-2 from the culture medium.
Claim 21
Dependent Claim
20 words
The process of claim 18, wherein said cloned malignant murine T-cells are cloned from T leukemic cells or lymphoma cells.
Claim 22
Dependent Claim
18 words
The process of claim 19, wherein said cloned T lymphoma cells are clones of LBRM-33 lymphoma murine T-cells.
Claim 23
Dependent Claim
31 words
The process of claim 18, 19 or 20 wherein the T-cell mitogen in the culture medium is a compound selected from the group consisting of phytohemagglutinin, concanavalin A, or pokeweed mitogen.
Claim 24
Dependent Claim
15 words
The process of claim 21, wherein the culture medium further includes a phorbol ester derivative.
Claim 25
Dependent Claim
15 words
The process of claim 22, wherein the phorbol ester derivative includes a phorbol myristate acetate.
Claim 26
Dependent Claim
27 words
The process of claims 18, 19, or 20, wherein the culture medium further includes either Roswell Park Memorial Institute medium, Click's medium or Dulbecco Modified Eagle medium.
Claim 27
Dependent Claim
25 words
The process of claims 18, 19 or 20, comprising recovering of the IL-2 from the culture medium after approximately sixteen to forty-eight hours of culturing.
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1986-10-24 | 1987-04-25 | 1987-10-25 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1990-10-24 | 1991-04-25 | 1991-10-25 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1994-10-24 | 1995-04-25 | 1995-10-25 | $7700.00 | Window Closed / Lapsed |