US4522918A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Process for producing monoclonal antibodies reactive with human breast cancer
Assignee: HEALTH AND HUMAN SERVICES United States, Secretary of, Department of
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Filing: 1981-12-15
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Granted: 1985-06-11
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Expires: 2001-12-15
Abstract
Abstract Monoclonal antibodies demonstrating a reactivity with human breast cancer are produced. The hybridoma cultures secreting immunoglobins are produced by hydridoma technology. Splenic lymphocytes of mice, immunized with membrane-enriched fractions of metastatic human mammary carcinoma tissue are fused with the NS-1 non-immunoglobulin-secreting murine myeloma cell line. Screening of immunoglobulin reactivities and double cloning of cultures yielded 11 monoclonal antibodies that demonstrated activities with the surface of human mammary tumor cells and not with the surface of apparently normal human tissues. These monoclonal antibodies aid in the diagnosis, prognosis and treatment of human breast cancer.
Inventors
- 👤 Jeffery Schlom
- 👤 David Colcher
- 👤 Marianna Nuti
- 👤 Patricia H. Hand
- 👤 Faye Austin
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 02:38
CPC Classifications
A; A61; A61K; A61K51/00; A61K51/02; A61K51/04; A61K51/08; A61K51/10; A61K51/1045; A61K51/1051; C; C07; C07K; C07K16/00; C07K16/18; C07K16/28; C07K16/30; C07K16/3015; A61K2123/00; A61K38/00; G; G01; G01N; G01N2474/00; G01N2474/20; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/948; Y10S436/00; Y10S436/811; Y10S436/813
Claims Analysis
Total Claims: 9
Claim 1
Independent Claim
130 words
A process for producing antibodies from hyridoma cultures comprising the steps of (1) taking splenic lymphocytes of mice previously immunized with membrane-enriched fractions of immunoglobulin-depleted cancer cells; (2) fusing the lymphocytes with murine myeloma cell line, and (3) culturing the cell line in an in vitro culture medium or in vivo therefor to produce antibodies, which antibody: (a) reacts and binds with extracts from human metastatic mammary carcinoma cells from involved livers but not with liver cell extracts; (b) reacts and binds with at least one of the mammary carcinoma cell lines, BT-20, MCF-7, ZR-75-1, but not with lung, vulva epidermoid or oral epidermoid and not with rhabdomyosarcoma, fibrosarcoma and melanoma; and (c) does not react with normal cells derived from breast, skin, lung, bone marrow, kidney, spleen and uterus.
Claim 2
Dependent Claim
130 words
A process for producing antibodies from hyridoma cultures comprising the steps of (1) taking splenic lymphocytes of mice previously immunized with membrane-enriched fractions of immunoglobulin-depleted cancer cells; (2) fusing the lymphocytes with murine myeloma cell line, and (3) culturing the cell line in an in vitro culture medium or in vivo therefor to produce antibodies, which antibody: (a) reacts and binds with extracts from human metastatic mammary carcinoma cells from involved livers but not with liver cell extracts; (b) reacts and binds with at least one of the mammary carcinoma cell lines, BT-20, MCF-7, ZR-75-1, but not with lung, vulva epidermoid or oral epidermoid and not with rhabdomyosarcoma, fibrosarcoma and melanoma; and (c) does not react with normal cells derived from breast, skin, lung, bone marrow, kidney, spleen and uterus.
Claim 3
Dependent Claim
19 words
The method according to claim 1 wherein said human cancer cells are from metastatic mammary carcinoma in the liver.
Claim 4
Dependent Claim
14 words
The method according to claim 1 wherein the depletion is performed using affinity chromatography.
Claim 5
Dependent Claim
19 words
The method according to claim 1 wherein the depletion is performed using protein A bound to a solid matrix.
Claim 6
Dependent Claim
11 words
The method according to claim 4 wherein said matrix is Sepharose.
Claim 7
Dependent Claim
21 words
A process of claim 1 wherein the murine myeloma cell line is non-immunoglobulin secreting murine myeloma cell line designated as NS-1.
Claim 8
Independent Claim
186 words
A method for preparing monoclonal antibody which antibody: (a) reacts and binds with extracts from human metastatic mammary carcinoma cells from involved livers but not with liver cell extracts; (b) reacts and binds with at least one of the mammary carcinoma cell lines, BT-20, MCF-7, ZR-75-1, but not with lung, vulva epidermoid or oral epidermoid and not with rhabdomyosarcoma, fibrosarcoma and melanoma; and (c) does not react with normal cells derived from breast, skin, lung, bone marrow, kidney, spleen and uterus, which comprises the steps of: (1) immunizing mice with membrane enriched fractions of immunoglobulin-depleted cancer cells; (2) removing the spleens from said mice and making a suspension of spleen cells; (3) fusing said spleen cells with NS-1 non-immunoglobulin-secreting murine myeloma cell line in the presence of a fusion promoter; (4) diluting and culturing the fused cells in separate wells in a medium which will not support the unfused cells; (5) evaluating the supernatant in each well containing a hybridoma for the presence of the desired antibody; (6) selecting and cloning hybridoma producing the desired antibody; and (7) recovering the antibody from supernatant above said clones.
Claim 9
Independent Claim
186 words
A method for preparing monoclonal antibody which antibody: (a) reacts and binds with extracts from human metastatic mammary carcinoma cells from involved livers but not with liver cell extracts; (b) reacts and binds with at least one of the mammary carcinoma cell lines, BT-20, MCF-7, ZR-75-1, but not with lung, vulva epidermoid or oral epidermoid and not with rhabdomyosarcoma, fibrosarcoma and melanoma; and (c) does not react with normal cells derived from breast, skin, lung, bone marrow, kidney, spleen and uterus, which comprises the steps of: (1) immunizing mice with membrane enriched fractions of immunoglobulin-depleted cancer cells; (2) removing the spleens from said mice and making a suspension of spleen cells; (3) fusing said spleen cells with NS-1 non-immunoglobulin-secreting murine myeloma cell line in the presence of a fusion promoter; (4) diluting and culturing the fused cells in separate wells in a medium which will not support the unfused cells; (5) evaluating the supernatant in each well containing a hybridoma for the presence of the desired antibody; (6) selecting and cloning hybridoma producing the desired antibody; and (7) recovering the antibody from supernatant above said clones.
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) | 1988-06-10 | 1988-12-10 | 1989-06-11 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1992-06-10 | 1992-12-10 | 1993-06-11 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1996-06-10 | 1996-12-10 | 1997-06-11 | $7700.00 | Window Closed / Lapsed |