EP0252685A2
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EP
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Diagnostic methods for detecting lymphomas in humans
Assignee: The Wistar Institute
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Filing: 1987-07-02
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Granted: —
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Expires: 2007-07-02
Abstract
Abstract The sequence of the protein coding regions of the bcl-2 gene are provided as well as bacterial clones which produce the proteins. Assays are provided for detecting a class of B-cell neoplasms associated with a chromosome translocation between chromosomes 14 and 18. This translocation is involved in the majority of cases of human follicular lymphomas. One assay employs an antibody which is immunoreactive with a human protein which is over-expressed due to the chromosome translocation. Another assay involves measurement of the amount of mRNA which hybridizes to the gene proximal to the translocation break-point.
Inventors
- 👤 Yoshihide Tsujimoto
- 👤 Carlos M Croce
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-05 23:08
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/82; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6876; C12Q1/6883; C12Q1/6886; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/575; G01N33/57575; C12Q2600/00; C12Q2600/156; C12Q2600/158; Y; Y10; Y10S; Y10S436/00; Y10S436/811; Y10S436/813; Y10S530/00; Y10S530/806; Y10S530/827; Y10S530/828
Claims Analysis
Total Claims: 21
Claim 1
Independent Claim
143 words
A diagnostic method for detecting B-cell neoplasms in a human, comprising the steps of: isolating B-cells from the human; extracting proteins from said B-cells to form a test sample; contacting said test sample with an antibody which is immunoreactive with a gene product of the bcl-2 gene under conditions where antibody-antigen complexes form and are stable to form antibody-antigen complexes; quantitating the amount of antibody-antigen complexes formed with said test sample; comparing the amount of antibody-antigen complexes formed with said test sample to the amount formed with a control sample of proteins, a ratio of test sample complexes to control sample complexes of greater than about ten indicating B-cell neoplasm, said control sample of proteins extracted from cells selected from the group consisting of: B-cells from a normal human, cells from an established normal B-cell or pre-B-cell line, and non-B-cells from said human.
Claim 2
Dependent Claim
143 words
A diagnostic method for detecting B-cell neoplasms in a human, comprising the steps of: isolating B-cells from the human; extracting proteins from said B-cells to form a test sample; contacting said test sample with an antibody which is immunoreactive with a gene product of the bcl-2 gene under conditions where antibody-antigen complexes form and are stable to form antibody-antigen complexes; quantitating the amount of antibody-antigen complexes formed with said test sample; comparing the amount of antibody-antigen complexes formed with said test sample to the amount formed with a control sample of proteins, a ratio of test sample complexes to control sample complexes of greater than about ten indicating B-cell neoplasm, said control sample of proteins extracted from cells selected from the group consisting of: B-cells from a normal human, cells from an established normal B-cell or pre-B-cell line, and non-B-cells from said human.
Claim 3
Dependent Claim
25 words
The method of claim 1 wherein the antibody is radiolabeled and the step of quantitating occurs by measuring the amount of radioactivity in antibody-antigen complexes.
Claim 4
Dependent Claim
32 words
The method of claim 1 wherein the antibody is conjugated to a fluorescent molecule and the step of quantitating is performed by measuring the amount of fluorescence emitted by said antibody-antigen complexes.
Claim 5
Dependent Claim
36 words
4 . The method of claim 1 wherein said step of quantitating is performed by means of use of a second antibody immunoreactive with said antibody which is immunoreactive with a gene product of bcl-2 gene.
Claim 6
Dependent Claim
12 words
The method of claim 1 wherein the antibody is immunoreactive with bcl-2-alpha.
Claim 7
Dependent Claim
12 words
The method of claim 1 wherein the antibody is immunoreactive with bcl-2-beta.
Claim 8
Dependent Claim
15 words
The method of claim 1 wherein the antibody is immunoreactive with both bel-2-alpha and bel-2-beta.
Claim 9
Dependent Claim
8 words
A human bcl-2 gene substantially free of introns.
Claim 10
Dependent Claim
8 words
A human bcl-2 gene substantially free of introns.
Claim 11
Dependent Claim
12 words
The gene of claim 8 wherein said gene is replicated in bacteria.
Claim 12
Dependent Claim
14 words
1 0. The gene of claim 9 wherein said gene is expressed in bacteria.
Claim 13
Dependent Claim
23 words
1 1. A substantially pure preparation of a protein having an N-terminal end encoded by the first exon of the human bcl-2 gene.
Claim 14
Dependent Claim
23 words
1 1. A substantially pure preparation of a protein having an N-terminal end encoded by the first exon of the human bcl-2 gene.
Claim 15
Dependent Claim
19 words
12 . The protein preparation of claim 11 wherein said protein is bcl-2-alpha having about 239 amino acid residues.
Claim 16
Dependent Claim
17 words
The protein preparation of claim 11 wherein said protein is bcl-2-beta having about 205 amino acid residues.
Claim 17
Dependent Claim
146 words
A diagnostic method for detecting B-cell neoplasms in a human comprising the steps of: isolating B-cells from the human; extracting RNA from said B-cells to form a test sample; contacting said test sample with a DNA probe containing a sequence of at least 15 nucleotides in length derived from the human bcl-2 gene under conditions where homolgous RNA-DNA hybrids form and are stable; quantitating the amount of RNA-DNA hybrids formed in the test sample; and comparing the amount of RNA-DNA hybrids formed with said test sample to the amount formed with a control sample of RNA, a ratio of test sample hybrids to control sample hybrids of greater than about ten indicating B-cell neoplasm, said control sample of RNA extracted from cells selected from the group consisting of: B-cells from a normal human, cells from a normal B-cell or pre-B-cell line, and non-B-cells from said human.
Claim 18
Dependent Claim
146 words
A diagnostic method for detecting B-cell neoplasms in a human comprising the steps of: isolating B-cells from the human; extracting RNA from said B-cells to form a test sample; contacting said test sample with a DNA probe containing a sequence of at least 15 nucleotides in length derived from the human bcl-2 gene under conditions where homolgous RNA-DNA hybrids form and are stable; quantitating the amount of RNA-DNA hybrids formed in the test sample; and comparing the amount of RNA-DNA hybrids formed with said test sample to the amount formed with a control sample of RNA, a ratio of test sample hybrids to control sample hybrids of greater than about ten indicating B-cell neoplasm, said control sample of RNA extracted from cells selected from the group consisting of: B-cells from a normal human, cells from a normal B-cell or pre-B-cell line, and non-B-cells from said human.
Claim 19
Dependent Claim
25 words
1 5. The method of claim 14 wherein said DNA probe hybridizes with the 8.5 kb, the 5.5 kb, and the 3.5 kb bcl-2 transcript.
Claim 20
Dependent Claim
20 words
The method of claim 14 wherein said DNA probe hybridizes with the 8.5 kb and the 5.5 kb bcl-2 transcript.
Claim 21
Dependent Claim
17 words
The method of. claim 14 wherein said DNA probe hybridizes exclusively with the 3.5 kb bcl-2 transcript.
References Cited (Backward Citations)
No citation records indexed.
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