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Patent US4444887A successfully refreshed from Google Patents.
US4444887A
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Process for making human antibody producing B-lymphocytes
Assignee: Memorial Sloan Kettering Cancer Center
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Filing: 1981-03-12
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Granted: 1984-04-24
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Expires: 2001-03-12
Abstract
Abstract Antibody producing human B-lymphocytes are produced by a process which comprises culturing human B-lymphocytes in a tissue culture medium in the presence of an antigen; helper signal producing agents comprising monocytes or monocyte conditioned medium containing Interleukin 1 (I1-1), and helper T-lymphocytes or helper T-lymphocyte replacing factor; and human serum; and thereafter recovering the antibody producing cells from the medium.
Inventors
- 👤 Michael K. Hoffmann
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-24 22:34
CPC Classifications
C; C12; C12N; C12N5/00; C12N5/10; C12N5/12; C12N5/16; C12N5/163; C07; C07K; C07K16/00; C12N2501/00; C12N2501/20; C12N2501/23; C12N2502/00; C12N2502/11
Claims Analysis
Total Claims: 30
Claim 1
Independent Claim
103 words
Process for making antibody producing human B-lymphocytes which comprises culturing a human lymphoid cell suspension containing human B-lymphocytes and a cell concentration of about 1×10 6 to 5×10 6 cells/ml in a tissue culture medium containing: (a) 0.3 to 0.003% antigen; (b) helper signal producing agents consisting essentially of: (i) monocytes or conditioned medium containing interleukin 1 derived from said monocytes in an amount to result in a concentration of 0.1 to 30% and (ii) helper T-lymphocytes or helper T-lymphocyte replacing factor obtained from activated T-cells; and (c) about 1-10% human serum; and thereafter recovering the antibody producing human B-Lymphocytes from the medium.
Claim 2
Dependent Claim
103 words
Process for making antibody producing human B-lymphocytes which comprises culturing a human lymphoid cell suspension containing human B-lymphocytes and a cell concentration of about 1×10 6 to 5×10 6 cells/ml in a tissue culture medium containing: (a) 0.3 to 0.003% antigen; (b) helper signal producing agents consisting essentially of: (i) monocytes or conditioned medium containing interleukin 1 derived from said monocytes in an amount to result in a concentration of 0.1 to 30% and (ii) helper T-lymphocytes or helper T-lymphocyte replacing factor obtained from activated T-cells; and (c) about 1-10% human serum; and thereafter recovering the antibody producing human B-Lymphocytes from the medium.
Claim 3
Dependent Claim
27 words
The process of claim 1 comprising the additional step of treating the lymphoid cell suspension to remove suppressor cells or their precursors prior to the culturing step.
Claim 4
Dependent Claim
27 words
The process of claim 1 wherein said tissue culture medium further comprises a B-lymphocyte mitogen at a concentration of about 0.3 to 0.00003%, as a proliferation promoter.
Claim 5
Dependent Claim
32 words
The process of claim 1 wherein said human lymphoid cell suspension contains in addition helper T-lymphocytes and said tissue culture medium comprises a T-lymphocyte mitogen as an activator for the helper T-lymphocytes.
Claim 6
Dependent Claim
23 words
The process of claim 1 comprising the additional step of treating the lymphoid cell suspension to remove T-lymphocytes prior to the culturing step.
Claim 7
Dependent Claim
25 words
Process of claim 5 wherein said removed T-lymphocytes are cultivated to produce T-lymphocyte helper factor which is isolated and added to the tissue culture medium.
Claim 8
Dependent Claim
11 words
Process of claim 1 wherein red blood cells comprise the antigen.
Claim 9
Dependent Claim
15 words
Process of claim 7 wherein the red blood cells are conjugated with a specific antigen.
Claim 10
Dependent Claim
13 words
Process of claim 1 wherein the antigen is concentration about 0.03%, by volume.
Claim 11
Dependent Claim
11 words
Process of claim 4 wherein the T-lymphocyte mitogen is Concanavalin A.
Claim 12
Dependent Claim
20 words
Process of claim 3 wherein the B-lymphocyte mitogen is the bacterial mitogen Staphylococcus aureus of concentration about 0.003%, by volume.
Claim 13
Dependent Claim
25 words
Process of claim 1 wherein said conditioned medium containing Interleukin 1 is in an amount to result in a concentration of about 10%, by volume.
Claim 14
Dependent Claim
15 words
Process of claim 1 wherein said human serum is a concentration about 5-10% by volume.
Claim 15
Dependent Claim
19 words
Process of claim 10 wherein said T-lymphocyte mitogen is Concanavalin A at a concentration of 0.3 to 10 μg/ml.
Claim 16
Dependent Claim
12 words
Process of claim 14 wherein said Concanavalin A is about 10 μg/ml.
Claim 17
Dependent Claim
12 words
Process of claim 1 wherein the culture medium is a Mishell-Dutton medium.
Claim 18
Dependent Claim
62 words
Process of claim 1 wherein the human lymphoid cell suspension contains in addition suppressor cells or their precursors and further comprises the steps of: (a) delaying the addition of human serum to the medium until 16 to 24 hours after culturing has started; and (b) adding to the culture medium initially 0.3 to 0.0003% of a B-lymphocyte mitogen as a proliferation promoter.
Claim 19
Dependent Claim
31 words
Process of claim 17 wherein said human lymphoid cell suspension contains in addition helper T-lymphocytes, further comprising the step of adding a T-lymphocyte mitogen as an activator for the helper T-lymphocytes.
Claim 20
Dependent Claim
17 words
Process of claim 17 comprising the additional step of removing T-lymphocytes from the cell suspension before culturing.
Claim 21
Dependent Claim
25 words
Process of claim 19 wherein the removed T-lymphocytes are cultivated to produce T-lymphocyte helper factor which is isolated and added to the B-lymphocyte culture medium.
Claim 22
Dependent Claim
11 words
Process of claim 17 wherein red blood cells comprise the antigen.
Claim 23
Dependent Claim
15 words
Process of claim 21 wherein the red blood cells are conjugated with a specific antigen.
Claim 24
Dependent Claim
19 words
Process of claim 17 wherein the antigen is added to the medium in concentrations of about 0.03% by volume.
Claim 25
Dependent Claim
13 words
Process of claim 17 wherein the mitogen is the bacterial mitogen, Staphylococcus aureus.
Claim 26
Dependent Claim
20 words
Process of claim 17 wherein the proliferation promoter is added to the medium in concentrations of about 0.003%, by volume.
Claim 27
Dependent Claim
24 words
Process of claim 17 wherein said conditioned medium containing Interleukin 1 is added to the culture medium in concentrations of about 10% by volume.
Claim 28
Dependent Claim
21 words
Process of claim 17 wherein the human serum is added to the culture medium in concentrations of about 5-10% by volume.
Claim 29
Dependent Claim
17 words
Process of claim 18 wherein 0.3 to 10 μg/ml Concanavalin A is added as a T-lymphocyte activator.
Claim 30
Dependent Claim
19 words
Process of claim 28 wherein the Concanavalin A is added to the medium in concentration of about 10 μg/ml.
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) | 1987-04-24 | 1987-10-24 | 1988-04-24 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1991-04-24 | 1991-10-24 | 1992-04-24 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1995-04-24 | 1995-10-24 | 1996-04-24 | $7700.00 | Window Closed / Lapsed |