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US4364934A Expired / Lapsed US ⏱️ 0.0 years remaining

Monoclonal antibody to a human early thymocyte antigen and methods for preparing same

Assignee: Ortho Pharmaceutical Corp
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Filing: 1979-12-04
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Granted: 1982-12-21
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Expires: 1999-12-04
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Abstract

Abstract Hybrid cell line for production of monoclonal antibody to an antigen found on approximately 10% of normal human thymocytes. The hybrid is formed by fusing splenocytes from immunized CAF 1 mice with P3X63Ag8U1 myeloma cells. Diagnostic and therapeutic uses of the monoclonal antibody are also disclosed.

Inventors

  • 👤 Patrick C. Kung
  • 👤 Gideon Goldstein

Bibliographic Details

Application Number —
Jurisdiction US
Data Source google_patents
Last Synchronized 2026-10-23 07:47
CPC Classifications
C; C07; C07K; C07K16/00; C07K16/18; C07K16/28; C07K16/2881; A; A61; A61P; A61P37/00; A61K; A61K38/00; Y; Y10; Y10S; Y10S530/00; Y10S530/808

Claims Analysis

Total Claims: 18
Claim 1
Independent Claim 205 words
A monoclonal antibody of class IgG produced by a hybridoma formed by fusion of cells from mouse myeloma line and spleen cells from a mouse previously immunized with human leukemic T-ALL cells, which antibody: (a) reacts with approximately 10 percent of normal human thymocytes, but not with normal human peripheral T cells, B cells, Null cells, or bone marrow cells, (b) reacts with 8 percent of untreated thymocytes, 6 percent of thymocytes previously treated with monoclonal antibody produced by hybrid cell line ATCC CRL 8002 and complement, 12 percent of thymocytes previously treated with monoclonal antibody produced by hybrid cell line ATCC CRL 8001 and complement, and 5 percent of the thymocytes previously treated with monoclonal antibody produced by hybrid cell line ATCC CRL 8014 and complement; (c) reacts with Early Thymocyte (Thy 2) Stage I and Common Thymocyte (Thy 3) Stage II T cell ALL Stages but not with Prothymocyte (Thy 1) Stage I, Common Thymocyte (Thy 4, Thy 3-4, or Thy 4-6) Stage II or Late Thymocyte (Thy 8) Stage III T cell ALL Stages; (d) reacts with HSB-2, CEM and MOLT-4 cell lines; and (e) defines a T cell antigen which appears on 30-50 percent of peripheral T cells after mitogen stimulation.
Claim 2
Dependent Claim 205 words
A monoclonal antibody of class IgG produced by a hybridoma formed by fusion of cells from mouse myeloma line and spleen cells from a mouse previously immunized with human leukemic T-ALL cells, which antibody: (a) reacts with approximately 10 percent of normal human thymocytes, but not with normal human peripheral T cells, B cells, Null cells, or bone marrow cells, (b) reacts with 8 percent of untreated thymocytes, 6 percent of thymocytes previously treated with monoclonal antibody produced by hybrid cell line ATCC CRL 8002 and complement, 12 percent of thymocytes previously treated with monoclonal antibody produced by hybrid cell line ATCC CRL 8001 and complement, and 5 percent of the thymocytes previously treated with monoclonal antibody produced by hybrid cell line ATCC CRL 8014 and complement; (c) reacts with Early Thymocyte (Thy 2) Stage I and Common Thymocyte (Thy 3) Stage II T cell ALL Stages but not with Prothymocyte (Thy 1) Stage I, Common Thymocyte (Thy 4, Thy 3-4, or Thy 4-6) Stage II or Late Thymocyte (Thy 8) Stage III T cell ALL Stages; (d) reacts with HSB-2, CEM and MOLT-4 cell lines; and (e) defines a T cell antigen which appears on 30-50 percent of peripheral T cells after mitogen stimulation.
Claim 3
Dependent Claim 13 words
The monoclonal antibody of claim 1 which is of subclass IgG 1 .
Claim 4
Dependent Claim 34 words
The monoclonal antibody of claim 1 which is produced from a hybridoma formed by fusion of P3X63Ag8U1 myeloma cells and spleen cells from a CAF 1 mouse previously immunized with human leukemic T-ALL cells.
Claim 5
Dependent Claim 59 words
A monoclonal antibody of class IgG produced by a hybridoma formed by fusion of cells from a mouse myeloma line and spleen cells from a mouse previously immunized with human leukemic T-ALL cells which reacts with approximately 10 percent of normal human thymocytes, but not with normal human peripheral T cells, B cells, Null cells, or bone marrow cells.
Claim 6
Dependent Claim 59 words
A monoclonal antibody of class IgG produced by a hybridoma formed by fusion of cells from a mouse myeloma line and spleen cells from a mouse previously immunized with human leukemic T-ALL cells which reacts with approximately 10 percent of normal human thymocytes, but not with normal human peripheral T cells, B cells, Null cells, or bone marrow cells.
Claim 7
Dependent Claim 28 words
Mouse monoclonal antibody which reacts with approximately 10% of normal human thymocytes but not with normal human peripheral T cells, B cells, null cells, or bone marrow cells.
Claim 8
Dependent Claim 28 words
Mouse monoclonal antibody which reacts with approximately 10% of normal human thymocytes but not with normal human peripheral T cells, B cells, null cells, or bone marrow cells.
Claim 9
Dependent Claim 166 words
A monoclonal antibody which reacts with approximately 10% of normal human thymocytes but not with normal human peripheral T cells, B cells, null cells, or bone marrow cells, prepared by the method which comprises the steps of: (i) immunizing mice with human leukemic T-ALL cells; (ii) removing the spleens from said mice and making a suspension of the spleen cells; (iii) fusing said spleen cells with mouse myeloma cells in the presence of a fusion promoter; (iv) diluting and culturing the fused cells in separate wells in a medium which will not support the unfused myeloma cells; (v) evaluating the supernatant in each well containing a hybridoma for the presence of antibody to E rosette positive purified T cells or human thymocytes; (vi) selecting and cloning a hybridoma producing antibody which reacts with approximately 10% of normal human thymocytes but not with normal human peripheral T cells, B cells, null cells, or bone marrow cells; and (vii) recovering the antibody from the supernatant above said clones.
Claim 10
Dependent Claim 166 words
A monoclonal antibody which reacts with approximately 10% of normal human thymocytes but not with normal human peripheral T cells, B cells, null cells, or bone marrow cells, prepared by the method which comprises the steps of: (i) immunizing mice with human leukemic T-ALL cells; (ii) removing the spleens from said mice and making a suspension of the spleen cells; (iii) fusing said spleen cells with mouse myeloma cells in the presence of a fusion promoter; (iv) diluting and culturing the fused cells in separate wells in a medium which will not support the unfused myeloma cells; (v) evaluating the supernatant in each well containing a hybridoma for the presence of antibody to E rosette positive purified T cells or human thymocytes; (vi) selecting and cloning a hybridoma producing antibody which reacts with approximately 10% of normal human thymocytes but not with normal human peripheral T cells, B cells, null cells, or bone marrow cells; and (vii) recovering the antibody from the supernatant above said clones.
Claim 11
Dependent Claim 181 words
A monoclonal antibody which reacts with approximately 10% of normal human thymocytes but not with normal human peripheral T cells, B cells, null cells, or bone marrow cells, prepared by the method, which comprises the steps of: (i) immunizing mice with human leukemic T-ALL cells; (ii) removing the spleens from said mice and making a suspension of the spleen cells; (iii) fusing said spleen cells with mouse myeloma cells in the presence of a fusion promoter; (iv) diluting and culturing the fused cells in separate wells in a medium which will not support the unfused myeloma cells; (v) evaluating the supernatant in each well containing a hybridoma for the presence of antibody to E rosette positive purified T cells or human thymocytes; (vi) selecting and cloning a hybridoma producing antibody which reacts with approximately 10% of normal human thymocytes but not with normal human peripheral T cells, B cells, null cells, or bone marrow cells; (vii) transferring said clones intraperitoneally into mice; and (viii) harvesting the malignant ascites or serum from said mice, which ascites or serum contains the desired antibody.
Claim 12
Dependent Claim 181 words
A monoclonal antibody which reacts with approximately 10% of normal human thymocytes but not with normal human peripheral T cells, B cells, null cells, or bone marrow cells, prepared by the method, which comprises the steps of: (i) immunizing mice with human leukemic T-ALL cells; (ii) removing the spleens from said mice and making a suspension of the spleen cells; (iii) fusing said spleen cells with mouse myeloma cells in the presence of a fusion promoter; (iv) diluting and culturing the fused cells in separate wells in a medium which will not support the unfused myeloma cells; (v) evaluating the supernatant in each well containing a hybridoma for the presence of antibody to E rosette positive purified T cells or human thymocytes; (vi) selecting and cloning a hybridoma producing antibody which reacts with approximately 10% of normal human thymocytes but not with normal human peripheral T cells, B cells, null cells, or bone marrow cells; (vii) transferring said clones intraperitoneally into mice; and (viii) harvesting the malignant ascites or serum from said mice, which ascites or serum contains the desired antibody.
Claim 13
Independent Claim 53 words
A method of preparing monoclonal antibody which reacts with approximately 10% of normal human thymocytes but not with normal human peripheral T cells, B cells, null cells, or bone marrow cells, which comprises culturing the hybridoma ATCC CRL 8021 in a suitable medium and recovering the antibody from the supernatant above said hybridoma.
Claim 14
Independent Claim 53 words
A method of preparing monoclonal antibody which reacts with approximately 10% of normal human thymocytes but not with normal human peripheral T cells, B cells, null cells, or bone marrow cells, which comprises culturing the hybridoma ATCC CRL 8021 in a suitable medium and recovering the antibody from the supernatant above said hybridoma.
Claim 15
Dependent Claim 10 words
The monoclonal antibody prepared by the method of claim 8.
Claim 16
Independent Claim 55 words
A method of preparing monoclonal antibody which reacts with approximately 10% of normal human thymocytes but not with normal human peripheral T cells, B cells, null cells, or bone marrow cells, which comprises injecting into a mouse the hybridoma ATCC CRL 8021 and recovering the antibody from the malignant ascites or serum of said mouse.
Claim 17
Independent Claim 55 words
A method of preparing monoclonal antibody which reacts with approximately 10% of normal human thymocytes but not with normal human peripheral T cells, B cells, null cells, or bone marrow cells, which comprises injecting into a mouse the hybridoma ATCC CRL 8021 and recovering the antibody from the malignant ascites or serum of said mouse.
Claim 18
Dependent Claim 10 words
The monoclonal antibody prepared by the method of claim 10.

References Cited (Backward Citations)

US4196265A Cited
US4172124A Cited
US4284412A Cited

USPTO Annuity & Maintenance Windows

Annuity Stage Window Opens Payment Deadline Surcharge Period Until Estimated Fee Status
3.5 Years (Year 4) 1985-12-20 1986-06-21 1986-12-21 $2000.00 Window Closed / Lapsed
7.5 Years (Year 8) 1989-12-20 1990-06-21 1990-12-21 $3760.00 Window Closed / Lapsed
11.5 Years (Year 12) 1993-12-20 1994-06-21 1994-12-21 $7700.00 Window Closed / Lapsed

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