US4876189A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Process for the quantification of cell populations or subpopulations and a reagent suitable therefor
Assignee: Roche Diagnostics GmbH
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Filing: 1986-11-05
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Granted: 1989-10-24
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Expires: 2006-11-05
Abstract
Abstract A process for quantification of cell populations or subpopulations, by incubating a sample with labelled antibodies directed against characteristic surface antigens of the cell population to be quantified to form labelled antibody/antigen complexes. Standards with known, differing particle concentration and having comparable sedimentation behavior to the cells to be determined and further, carrying molecules which are directed against the labelled antibody or a part hereof are also incubated with the labelled antibodies. The cells of the sample solution, as well as the particles of the standard solution, are separated off from the excess labelled antibodies and the amount of the labelling is measured not only on the cells but also on the particles. By comparison of the measurement value from the sample with the measurement values from the standard solutions, there is ascertained the number of cells to be determined in the sample. The standard and a reagent kit for carrying out the process are also the subject of this application.
Inventors
- 👤 Hartmut Schetters
- 👤 Josef Endl
- 👤 Winfried Albert
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-02 18:23
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/536; G01N33/537; G01N33/569; G01N33/56966; G01N33/56972; Y; Y10; Y10S; Y10S435/00; Y10S435/81; Y10S435/967; Y10T; Y10T436/00; Y10T436/10; Y10T436/101666
Claims Analysis
Total Claims: 12
Claim 1
Independent Claim
265 words
A process for the determination of the number of cells in a cell population or a cell subpopulation in a sample, comprising: (1) providing a sample of cells; (2) incubating said sample of cells with labelled antibodies specific to cell surface antigens of a cell population or a cell subpopulation under conditions favorable to a reaction between said cell surface antigens and said labelled antibodies, whereby the labelled antibodies complex with the cell surface antigens; (3) providing a standard mixture of particles having a known particle concentration, said standard mixture of particles having a sedimentation behavior similar to the cells to be determined, a first portion of said mixture of particles comprising particles carrying a non-specific antibody and a second portion of said mixture of particles carrying molecules directed specifically against the labeled antibodies or a part thereof; (4) incubating said standard mixture of particles with another portion of the labeled antibodies under conditions similar to conditions as the incubation of said sample in step (2), whereby the labeled antibodies complex with said second portions of said mixture of particles; (5) separating the cells from the labeled antibodies that do not complex with the cell surface antigens; (6) separating the particles from labeled antibodies that do not complex with the second portion of the mixture of particles; (7) measuring the amount of labeling present on the separated cells and present on the separated particles; and (8) determining the number of cells by comparing a value obtained by measuring the label on said cells with value obtained by measuring label attached to said mixture of particles.
Claim 2
Dependent Claim
265 words
A process for the determination of the number of cells in a cell population or a cell subpopulation in a sample, comprising: (1) providing a sample of cells; (2) incubating said sample of cells with labelled antibodies specific to cell surface antigens of a cell population or a cell subpopulation under conditions favorable to a reaction between said cell surface antigens and said labelled antibodies, whereby the labelled antibodies complex with the cell surface antigens; (3) providing a standard mixture of particles having a known particle concentration, said standard mixture of particles having a sedimentation behavior similar to the cells to be determined, a first portion of said mixture of particles comprising particles carrying a non-specific antibody and a second portion of said mixture of particles carrying molecules directed specifically against the labeled antibodies or a part thereof; (4) incubating said standard mixture of particles with another portion of the labeled antibodies under conditions similar to conditions as the incubation of said sample in step (2), whereby the labeled antibodies complex with said second portions of said mixture of particles; (5) separating the cells from the labeled antibodies that do not complex with the cell surface antigens; (6) separating the particles from labeled antibodies that do not complex with the second portion of the mixture of particles; (7) measuring the amount of labeling present on the separated cells and present on the separated particles; and (8) determining the number of cells by comparing a value obtained by measuring the label on said cells with value obtained by measuring label attached to said mixture of particles.
Claim 3
Dependent Claim
134 words
The process of claim 1 further comprising: (9) providing a second standard mixture of particles having a known particle concentration and a sedimentation behavior similar to the cells to be determined, and having said first and second portions thereof mixed in a different defined proportion from that of the first standard mixture; (10) incubating said second standard mixture of particles with another portion of labeled antibodies under conditions similar to conditions as the incubation of said sample in step (2); (11) separating the particles from the labeled antibodies that do not complex with the second portion of the second mixture of particules; (12) measuring the amount of labeling present on the separated particles obtained in step (11); (13) employing the measured amount obtained in step (12) in the comparing of values in step (8).
Claim 4
Dependent Claim
13 words
The process of claim 1, wherein said labelled antibodies are labelled monoclonal antibodies.
Claim 5
Dependent Claim
18 words
The process of claim 1, wherein the label on the antibodies is a radioisotope, a dyestuff or enzyme.
Claim 6
Dependent Claim
12 words
The process of claim 4, wherein said dyestuff is a fluorescent dyestuff.
Claim 7
Dependent Claim
19 words
The process of claim 1, wherein the label on the antibodies is peroxidase, alkaline phosphatase, glucose oxidase or beta-galactosidase.
Claim 8
Dependent Claim
15 words
The process of claim 1, wherein said antibodies are antibodies against surface antigens of T-lymphocytes.
Claim 9
Dependent Claim
17 words
The process of claim 1, wherein said antibodies are antibodies against surface antigen of T-helper cell subpopulations.
Claim 10
Dependent Claim
15 words
The process of claim 1, wherein said labeled antibodies are antibodies against T-suppressor cell subpopulations.
Claim 11
Dependent Claim
95 words
A reagent kit for carrying out the process of claim 1 comprising: a labelled antibody which is specifically directed against the cell population or subpopulation to be determined; one or more standards comprising a mixture of particles which mixture has a sedimentation behavior similar to the population or subpopulation to be determined, a first portion of said mixture of particles carrying a non-specific antibody and a second portion of said mixture of particles carrying molecules specifically directed against said labeled antibodies or a part thereof; and a reagent for the measurement of the antibody label.
Claim 12
Dependent Claim
22 words
The reagent kit of claim 10, wherein the antibody is labelled with beta-galactosidase and the reagent is for the detection of beta-galactosidase.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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| 3.5 Years (Year 4) | 1992-10-23 | 1993-04-24 | 1993-10-24 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1996-10-23 | 1997-04-24 | 1997-10-24 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2000-10-23 | 2001-04-24 | 2001-10-24 | $7700.00 | Window Closed / Lapsed |