US4448890A
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Detection of human cancer cells with antibodies to human cancer nucleolar antigens
Assignee: Baylor College of Medicine
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Filing: 1980-09-29
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Granted: 1984-05-15
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Expires: 2000-09-29
Abstract
Abstract Specific common nucleolar antigens are found in a broad range of human malignant tumor specimens and have been isolated, extracted and purified. Antibodies and antisera specific to these nucleolar antigens are harvested from non-human hosts and used for detection of human cancer cells. Disclosed are (1) methods for isolating the nucleolar antigens, obtaining them in substantially purified form, producing the specific antibodies, obtaining them in substantially purified form, and using the antisera and antibodies induced by the nucleolar antigens in diagnostic procedures for detecting human cancer cells, and (2) diagnostic kits comprising specific antibodies and antisera.
Inventors
- 👤 Karel Smetana
- 👤 Harris Busch
- 👤 Rose K. Busch
- 👤 Ferenc Gyorkey
- 👤 Phyllis Gyorkey
- 👤 Frances M. Davis
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-26 23:08
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/575; G01N33/5758; G01N33/57595; Y; Y10; Y10S; Y10S436/00; Y10S436/807; Y10S436/808; Y10S436/811; Y10S436/813; Y10S530/00; Y10S530/806; Y10S530/827; Y10S530/828
Claims Analysis
Total Claims: 19
Claim 1
Independent Claim
76 words
Antigens selected from the group consisting of isolated nucleoli and nuclear extracts which contain nucleoli constituents, the antigens being in substantially purified form and having major and minor species, the major species having the following characteristics, a pI on isoelectric focusing of about 6.0 to 6.7, and a molecular weight of about 50,000 to about 60,000 daltons, is soluble in 0.01 M Tris HCl pH 8, and is primarily localized in nucleoli of human cancer cells.
Claim 2
Dependent Claim
76 words
Antigens selected from the group consisting of isolated nucleoli and nuclear extracts which contain nucleoli constituents, the antigens being in substantially purified form and having major and minor species, the major species having the following characteristics, a pI on isoelectric focusing of about 6.0 to 6.7, and a molecular weight of about 50,000 to about 60,000 daltons, is soluble in 0.01 M Tris HCl pH 8, and is primarily localized in nucleoli of human cancer cells.
Claim 3
Dependent Claim
96 words
Antigens selected from the group consisting of isolated nucleoli and nuclear extracts which contain nucleoli constituents, the antigens having major and minor species, the major species having the following characteristics, a pI on isoelectric focusing of about 6.0 to 6.7, and a molecular weight of about 50,000 to about 60,000 daltons, is soluble in 0.01 M Tris HCl pH 8, is primarily localized in nucleoli of human cancer cells, and is substantially purified by sequential purification by ammonium sulfate precipitation, DEAE-column chromatography, Sephadex gel exclusion, cation exchange and calcium phosphate gel chromatography, and preparative isoelectric focusing.
Claim 4
Dependent Claim
96 words
Antigens selected from the group consisting of isolated nucleoli and nuclear extracts which contain nucleoli constituents, the antigens having major and minor species, the major species having the following characteristics, a pI on isoelectric focusing of about 6.0 to 6.7, and a molecular weight of about 50,000 to about 60,000 daltons, is soluble in 0.01 M Tris HCl pH 8, is primarily localized in nucleoli of human cancer cells, and is substantially purified by sequential purification by ammonium sulfate precipitation, DEAE-column chromatography, Sephadex gel exclusion, cation exchange and calcium phosphate gel chromatography, and preparative isoelectric focusing.
Claim 5
Dependent Claim
97 words
A diagnostic kit comprising, antibodies characterized by specificity against human cell associated nucleolar antigens in substantially purified form having major and minor species, the major species having the following characteristics, a pI on isoelectric focusing of about 6.0 to 6.7, and a molecular weight of about 50,000 to about 60,000 daltons, is soluble in 0.01 M Tris HCl pH 8, and is primarily localized in nucleoli of human cancer cells, the antibodies having a label detectable by one of light, fluorescence, electron microscopy, indirect assay methods and autoradiography, and disposed in a buffered suspending agent or solution.
Claim 6
Dependent Claim
97 words
A diagnostic kit comprising, antibodies characterized by specificity against human cell associated nucleolar antigens in substantially purified form having major and minor species, the major species having the following characteristics, a pI on isoelectric focusing of about 6.0 to 6.7, and a molecular weight of about 50,000 to about 60,000 daltons, is soluble in 0.01 M Tris HCl pH 8, and is primarily localized in nucleoli of human cancer cells, the antibodies having a label detectable by one of light, fluorescence, electron microscopy, indirect assay methods and autoradiography, and disposed in a buffered suspending agent or solution.
Claim 7
Dependent Claim
132 words
A diagnostic kit containing, antibodies characterized by specificity against antigens selected from the group consisting of isolated nucleoli and nuclear extracts which contain nucleoli constituents, the antigens being in substantially purified form and having major an minor species, the major species having the following characteristics, a pI on isoelectric focusing of about 6.0 to 6.7, and a molecular weight of about 50,000 to about 60,000 daltons, is soluble in 0.01 M Tris HCI pH 8, and is primarily localized in nucleoli of human cancer cells, the primary antibodies being unlabeled and in a buffered suspended agent or solution, and appropriate labeled or unlabeled reagents for detection of the primary antibodies in fixed human malignant cells by a method selected from the group consisting of live micrscopy, fluorescent microscopy, electron microscopy and autoradiography.
Claim 8
Dependent Claim
132 words
A diagnostic kit containing, antibodies characterized by specificity against antigens selected from the group consisting of isolated nucleoli and nuclear extracts which contain nucleoli constituents, the antigens being in substantially purified form and having major an minor species, the major species having the following characteristics, a pI on isoelectric focusing of about 6.0 to 6.7, and a molecular weight of about 50,000 to about 60,000 daltons, is soluble in 0.01 M Tris HCI pH 8, and is primarily localized in nucleoli of human cancer cells, the primary antibodies being unlabeled and in a buffered suspended agent or solution, and appropriate labeled or unlabeled reagents for detection of the primary antibodies in fixed human malignant cells by a method selected from the group consisting of live micrscopy, fluorescent microscopy, electron microscopy and autoradiography.
Claim 9
Dependent Claim
45 words
A process for the immunological detection of cancer in human tissue sections, smears and exfoliative cytological preparations comprising, contacting specimens with the antibodies of claims 3 or 4, and demonstrating the localization of said antibodies in the nucleoli of malignant calles but not normal cells.
Claim 10
Dependent Claim
31 words
The process of claim 5 wherein, the antibodies are elicited to nucleoli isolated from one of the family of malignant human cells consisting of HeLa cells, carcinomas, sarcomas, and hematological neoplasms.
Claim 11
Dependent Claim
27 words
The process of claim 5 wherein, the antibodies are elicited to nucleoli isolated from one of the group consisting of HeLa cells or human prostatic carcinoma cells.
Claim 12
Dependent Claim
32 words
The process of claim 5 wherein, the antibodies are elicited to extracts of nuclei or nucleoli of malignant human cells from the family consisting of HeLa cells, carcinomas, sarcomas, and hematological neoplasms.
Claim 13
Dependent Claim
30 words
The process of claim 5 wherein, the antibodies are elicited to extracts of nuclei or nucleoli of one of the group consisting of HeLa cells or human prostatic carcinoma cells.
Claim 14
Dependent Claim
30 words
The process of claim 5 wherein, the antibodies are elicited to extracts of nuclei or nucleoli of malignant human cells obtained by extraction of 0.01 M Tris HCl, pH 7-9.
Claim 15
Dependent Claim
22 words
The process of claim 5 wherein, the immunological detection of cancer is by indirect immunochemical demonstration of the antibodies of claim 4.
Claim 16
Independent Claim
30 words
A method of purifying nucleolar antigens comprising, sequential purification thereof by ammonium sulfate precipitation, DEAE-column chromatography, Sephadex gel exclusion, cation exchange and calcium phosphate gel chromatography, and preparative isoelectric focusing.
Claim 17
Independent Claim
30 words
A method of purifying nucleolar antigens comprising, sequential purification thereof by ammonium sulfate precipitation, DEAE-column chromatography, Sephadex gel exclusion, cation exchange and calcium phosphate gel chromatography, and preparative isoelectric focusing.
Claim 18
Dependent Claim
19 words
The process of claim 12 where, the nucleolar antigens are extracted from nuclei or nucleoli from human cancer cells.
Claim 19
Dependent Claim
38 words
A process of preparing antibodies with specificity against antigens found in nucleoli of human cancer cells comprising, immunizing a non-human host animal with the antigens of claims 1 or 2, and harvesting the antibodies of the immunized animal.
References Cited (Backward Citations)
US4296201A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1987-05-15 | 1987-11-14 | 1988-05-15 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1991-05-15 | 1991-11-14 | 1992-05-15 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1995-05-15 | 1995-11-14 | 1996-05-15 | $7700.00 | Window Closed / Lapsed |