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US5068175A
Expired / Lapsed
US
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Method of detecting ras oncogene related malignancies
Assignee: The University of Texas System
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Filing: 1989-05-26
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Granted: 1991-11-26
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Expires: 2009-05-26
Abstract
Abstract Methods and composition for detecting the presence of human ras oncogene related malignancies are provided, where a biological sample is assayed for protein(s) specific to a DNA sequence. In the methods of this invention, the test reaction involves admixing a biological sample from cancer patients or control donors with labelled ras oncogene promoter DNA. The admixture is incubated under conditions favorable for promoting specific interactions between proteins and the labelled DNA. Thereafter, the admixture is separated by charge and size in an electrophoretic field and the protein-DNA interactions are identified depending on the method of label employed. Bands migrating at a slower rate than the uncomplexed DNA are indicative of a protein-DNA interaction (i.e. circulating serum protein(s) from cancer patients interacting specifically with a region(s) of the ras oncogene promoter DNA). Utilizing this experimental protocol, the serum proteins of interest include at least four different proteins that specifically interact with a region or regions of the ras oncogene promoter DNA. The four different factors, ranging in molecular weight from about 200 Kd to about 50 Kd are proteinaceous in nature as demonstrated by their trypsin sensitivity and heat stability.
Inventors
- 👤 Nagindra Prashad
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-06 05:09
CPC Classifications
C; 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; Y; Y10; Y10S; Y10S436/00; Y10S436/811; Y10S436/813
Claims Analysis
Total Claims: 19
Claim 1
Independent Claim
147 words
A method for detecting ras oncogene related malignancy in a human subject, comprising the steps of: a) collecting a test serum or plasma sample from a human subject; b) providing a control serum or plasma sample from a tumor free donor; c) incubating each of the control and test samples with a DNA having a sequence consisting essentially of a human ras oncogene promoter under conditions effective to promote specific binding interactions between protein factors in the samples and the DNA sequence; d) separating in each of the control and test samples a protein factor-DNA complex from free DNA; and e) comparing the amounts of protein factor-DNA complex of both the test and control samples, wherein if the test sample features an amount of about 10-fold or higher of a protein factor-DNA complex than the control sample, there is an indication of a ras oncogene related malignancy.
Claim 2
Independent Claim
147 words
A method for detecting ras oncogene related malignancy in a human subject, comprising the steps of: a) collecting a test serum or plasma sample from a human subject; b) providing a control serum or plasma sample from a tumor free donor; c) incubating each of the control and test samples with a DNA having a sequence consisting essentially of a human ras oncogene promoter under conditions effective to promote specific binding interactions between protein factors in the samples and the DNA sequence; d) separating in each of the control and test samples a protein factor-DNA complex from free DNA; and e) comparing the amounts of protein factor-DNA complex of both the test and control samples, wherein if the test sample features an amount of about 10-fold or higher of a protein factor-DNA complex than the control sample, there is an indication of a ras oncogene related malignancy.
Claim 3
Dependent Claim
14 words
The method of claim 1, wherein the ras oncogene related malignancy is prostate cancer.
Claim 4
Dependent Claim
14 words
The method of claim 1, wherein the ras oncogene related malignancy is breast cancer.
Claim 5
Dependent Claim
14 words
The method of claim 1, wherein the ras oncogene related malignancy is colon cancer.
Claim 6
Dependent Claim
13 words
The method of claim 1, wherein a ras oncogene related malignancy is melanoma.
Claim 7
Dependent Claim
30 words
The method according to claim 1, wherein the step of detecting ras oncogene related malignancy includes the step of separating the factor-DNA complex from the reaction mixture by gel electrophoresis.
Claim 8
Dependent Claim
27 words
The DNA of claim 1, wherein the human ras oncogene promoter is derived from a family of genes consisting of either Harvey, Kirsten or neuroblastoma ras oncogene.
Claim 9
Dependent Claim
34 words
The method of claim 1, wherein the human ras oncogene promoter is a human Harvey ras oncogene promoter comprising a Nae 1-Nae 1 DNA fragment encompassing 551 base pairs of nucleotide sequences from 116-666.
Claim 10
Dependent Claim
25 words
The method of claim 1, wherein the DNA consists essentially of an entire DNA sequence specific for the coding of the human Harvey ras oncogene.
Claim 11
Dependent Claim
29 words
The method of claim 1, wherein the DNA consists essentially of the ras promoter and a DNA sequence specific for the coding of a heterologous eukaryotic or prokaryotic gene.
Claim 12
Dependent Claim
27 words
The method of claim 1, wherein the protein factor-DNA complex comprises 4 different proteins each ranging in molecular weight from about 200 Kd to about 50 Kd.
Claim 13
Independent Claim
80 words
A method for diagnosing ras oncogene related malignancy in a human subject, the method comprising: detecting in a serum or plasma sample from the human subject, proteins complexed with a DNA having a sequence consisting essentially of a human ras oncogene promoter, the proteins having a range of molecular weights from about 200 Kd to about 50 Kd according to their relative electrophoretic migration rate through a gel matrix, and such complexes being an indicator of ras oncogene related malignancy.
Claim 14
Independent Claim
80 words
A method for diagnosing ras oncogene related malignancy in a human subject, the method comprising: detecting in a serum or plasma sample from the human subject, proteins complexed with a DNA having a sequence consisting essentially of a human ras oncogene promoter, the proteins having a range of molecular weights from about 200 Kd to about 50 Kd according to their relative electrophoretic migration rate through a gel matrix, and such complexes being an indicator of ras oncogene related malignancy.
Claim 15
Dependent Claim
15 words
A method according to claim 12, wherein the protein is trypsin sensitive and heat stable.
Claim 16
Dependent Claim
17 words
A method according to claim 12, wherein the labelled DNA comprises the human Harvey ras promoter oncogene.
Claim 17
Dependent Claim
76 words
A diagnostic kit for detecting an oncogene related malignancy in a human subject which comprises multiple containers wherein the following is included in separate containers: a human ras oncogene promoter DNA sequence, a positive control comprised of a protein which specifically binds to the ras oncogene promoter and a negative control comprised of a protein which does not bind to the promoter; the kit includes instructions which describe the separation and identification of a protein-DNA complex.
Claim 18
Dependent Claim
76 words
A diagnostic kit for detecting an oncogene related malignancy in a human subject which comprises multiple containers wherein the following is included in separate containers: a human ras oncogene promoter DNA sequence, a positive control comprised of a protein which specifically binds to the ras oncogene promoter and a negative control comprised of a protein which does not bind to the promoter; the kit includes instructions which describe the separation and identification of a protein-DNA complex.
Claim 19
Dependent Claim
50 words
The diagnostic kit of claim 15, wherein the ras oncogene promoter DNA comprises the promoter region alone, the promoter region added to a DNA sequence specific for the coding of an oncogene, or the promoter region added to a DNA sequence specific for coding any other eukaryotic or prokaryotic gene.
References Cited (Backward Citations)
US4699877A
Cited
US4963658A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1994-11-25 | 1995-05-27 | 1995-11-26 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1998-11-25 | 1999-05-27 | 1999-11-26 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2002-11-25 | 2003-05-27 | 2003-11-26 | $7700.00 | Window Closed / Lapsed |