EP0102228A2
Expired / Lapsed
EP
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Hybridoma secreting a monoclonal antibody specific for 5-bromo and 5-iododeoxyuridine and reagent for measuring cellular proliferation
Assignee: University of Miami
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Filing: 1983-08-19
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Granted: —
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Expires: 2003-08-19
Abstract
Abstract Cells synthesizing DNA are detected in a rapid, nonradioactive assay using the monoclonal antibody reagent secreted by the hybridoma produced by fusion of a myeloma cell, preferably murine plasmacytoma cell, with an animal antibody producing cell, preferably a mouse spleen cell, immunized with the conjugate of 5-bromouridine or 5-iodouridineand an albumin-carrier molecule. The antibody is specific for 5-bromodeoxyuridine or 5-iododeoxyuridine.
Inventors
- 👤 Howard George Gratzner
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-29 21:41
CPC Classifications
C; C07; C07K; C07K16/00; C07K16/44; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/5308; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/948
Claims Analysis
Total Claims: 18
Claim 1
Independent Claim
45 words
A hybridoma characterised in that it is a hybrid cell resulting from the fusion of a myeloma cell with an animal antibody-producing cell immunized with the conjugate of 5-bromouridine or 5-iodouridine and an albumin carrier molecule, said cell producing antibodies specific for 5-bromodeoxyuridine or 5-iododeoxyuridine.
Claim 2
Dependent Claim
45 words
A hybridoma characterised in that it is a hybrid cell resulting from the fusion of a myeloma cell with an animal antibody-producing cell immunized with the conjugate of 5-bromouridine or 5-iodouridine and an albumin carrier molecule, said cell producing antibodies specific for 5-bromodeoxyuridine or 5-iododeoxyuridine.
Claim 3
Dependent Claim
18 words
A hybridoma as claimed in Claim 1, characterised in that the animal antibody-producing cell is a spleen cell.
Claim 4
Dependent Claim
18 words
A hybridoma as claimed in Claim 2, characterised in that the spleen cell is a mouse spleen cell.
Claim 5
Dependent Claim
22 words
A hybridoma as claimed in any one of the preceding claims characterised in that the myeloma cell is a murine plasmacytoma cell.
Claim 6
Dependent Claim
23 words
A hybridoma as claimed in Claim 4, characterised in that the murine plasmacytoma cell is from plasmacytoma cell line SP 2/0 Ag 15,m.
Claim 7
Dependent Claim
24 words
A hybridoma as claimed in Claim 5, characterised in that it is deposited at the American Type Culture Collection under ATCC accession number HB-8150.
Claim 8
Dependent Claim
26 words
A composition comprising a hybridoma and a cell culture medium therefor, characterised in that the hybridoma is as claimed in any one of the preceding claims.
Claim 9
Dependent Claim
18 words
A composition as claimed in Claim 7, characterised in that the culture medium contains hypoxanthine- methotrexate-thymidine (HAT) medium.
Claim 10
Dependent Claim
32 words
A monoclonal antibody secreted by a hybridoma, characterised in that the hybridoma is as defined in any one of Claims 1 to 6 whereby the antibody is specific for 5-bromodeoxyuridine or 5-iododeoxyuridine.
Claim 11
Independent Claim
80 words
A method of producing a hybridoma as defined in any one of Claims 1 to 6, wherein an animal is immunized with a conjugate, an antibody-producing cell of the immunized animal is fused with a myeloma cell, the hybrids so produced are cloned, and clones producing antibodies demonstrating a specificity are selected characterised in that the conjugate is a conjugate of 5-bromouridine or 5-iodouridine and an albumin carrier molecule and the clones demonstrating specificity for 5-bromodeoxyuridine or 5-iododeoxyuridine are selected.
Claim 12
Dependent Claim
29 words
A method of producing antibodies as defined in Claim 9, characterised in that hybridomas are produced according to the method of Claim 7 and the antibodies are collected therefrom.
Claim 13
Dependent Claim
26 words
A reagent composition for detecting cellular proliferation via DNA synthesis, characterised in that it comprises, as the reagent-active ingredient, an antibody as defined in Claim 9.
Claim 14
Dependent Claim
23 words
A reagent composition as claimed in Claim 12, characterised in that the antibody has a fluorescent molecule detectable by immunofluorescent bound to it.
Claim 15
Dependent Claim
26 words
A reagent composition as claimed in Claim 12, characterised in that the antibody has a fluorescent enzyme detectable by enzyme linked immunoabsorbent assay bound to it.
Claim 16
Dependent Claim
48 words
A method of measuring the rate of DNA synthesis in a single cell by incorporating an antibody into DNA of the cell and thereafter measuring the antibody in the cellular DNA as an indication of DNA synthesis, characterised in that the antibody is as defined in Claim 9.
Claim 17
Dependent Claim
28 words
A method as claimed in Claim 15, characterised in that the said antibody has a fluorescent molecule bound to it and the said measurement is an immunofluorescent assay,
Claim 18
Dependent Claim
31 words
A method as claimed in Claim 15, characterised in that the said antibody has a fluorescent enzyme bound to it and the said measurement is by an enzyme linked immunoabsorbent assay.
References Cited (Backward Citations)
No citation records indexed.
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