Patent US5256532A successfully refreshed from Google Patents.
US5256532A
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US
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Methods, reagents and test kits for determination of subpopulations of biological entities
Assignee: CREDITANSTALT BANKVEREIN
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Filing: 1989-05-01
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Granted: 1993-10-26
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Expires: 2009-05-01
Abstract
Abstract Analytes, having a characteristic determinant that selectively interacts with a specific binding substance, are determined by coupling a reporter substance to the analyte, or to the specific binding substance, causing complex formation between the analyte and the specific binding substance in a test medium, separating the complexes thus formed from the test medium, and determining the presence or quantity of the analyte of interest by detecting the occurrence of the reporter substance in the complexes or the separated test medium. The determination is preferably performed on biomembrane-containing entities, such as cell subpopulations and/or subsets thereof, the reporter substance being stably associated with the lipid component of the biomembrane. Reagents and test kits are disclosed for performing the analyte determination.
Inventors
- 👤 Meryle J. Melnicoff
- 👤 Katharine A. Muirhead
- 👤 Paul K. Horan
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-08 09:15
CPC Classifications
A; A61; A61K; A61K41/00; A61K41/0085; A61K47/00; A61K47/50; A61K47/51; A61K47/56; A61K47/59; A61K47/60; A61K49/00; A61K49/0002; A61K49/001; A61K49/0013; A61K49/0017; A61K49/0019; A61K49/0021; A61K49/0032; A61K49/06; A61K49/18; A61K49/1896; C; C07; C07C; C07C229/00; C07C229/02; C07C229/04; C07C229/06; C07C229/10; C07C229/16; C07D; C07D209/00; C07D209/02; C07D209/04; C07D209/08; C07D219/00; C07D219/04; C07D219/08; C07D263/00; C07D263/52; C07D263/62; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/569; G01N33/56966; G01N33/56972; Y; Y10; Y10T; Y10T436/00; Y10T436/24
Claims Analysis
Total Claims: 61
Claim 1
Independent Claim
177 words
A method for determining the presence or quantity of analyte selected from the group consisting of eucaryotic cells, procaryotic cells and viruses, having lipid-containing membranes and at least one characteristic determinant, in a test sample suspected of containing said analyte, comprising the steps of: (i) coupling a detectable reproter substance non-selectively with said analyte in said test sample, said detectable reporter substance having a lipophilic moiety, which stably associates with said lip-containing membrane of said analyte; (ii) contacting said test sample with a specific binding substance which binds specifically to said at least one characteristic determinant of said analyte, under conditions causing complex formation between said analyte and said specific binding substance; (iii) after said coupling step and said contacting step, separating said test sample into a portion containing said complex and a portion substantially free of said complexs; and (iv) detecting the occurrence of said reporter substance in one of said separated portions to provide an indication of the presence or quantity of said analyte having said at least one characterstic determinant in said test sample.
Claim 2
Independent Claim
177 words
A method for determining the presence or quantity of analyte selected from the group consisting of eucaryotic cells, procaryotic cells and viruses, having lipid-containing membranes and at least one characteristic determinant, in a test sample suspected of containing said analyte, comprising the steps of: (i) coupling a detectable reproter substance non-selectively with said analyte in said test sample, said detectable reporter substance having a lipophilic moiety, which stably associates with said lip-containing membrane of said analyte; (ii) contacting said test sample with a specific binding substance which binds specifically to said at least one characteristic determinant of said analyte, under conditions causing complex formation between said analyte and said specific binding substance; (iii) after said coupling step and said contacting step, separating said test sample into a portion containing said complex and a portion substantially free of said complexs; and (iv) detecting the occurrence of said reporter substance in one of said separated portions to provide an indication of the presence or quantity of said analyte having said at least one characterstic determinant in said test sample.
Claim 3
Dependent Claim
36 words
A method according to claim 1, which also includes the step of relating the level of detected reporter substance to a predetermined standard to determine the presence or quantity of said analyte in said test sample.
Claim 4
Dependent Claim
43 words
A method according to claim 2, wherein the level of detected reporter substance is related to a standard containing a predetermined quantity of said reporter substance, thereby to quantiatively determine the analyte having said at least one characteristic determinant in said test sample.
Claim 5
Dependent Claim
20 words
A method according to claim 1, wherein the occurrence of said detectable reporter substance is detected in said complex-containing portion.
Claim 6
Dependent Claim
25 words
A method, according to claim 1, wherein said contacting step comprises contacting said test sample with said specific binding substance affixed to a solid phase.
Claim 7
Dependent Claim
35 words
A method according to claim 5, wherein the solid phase to which the specific binding substance is affixed comprises magnetic material, and said solid phase bearing said complex is magnetically separated from said complex-free portion.
Claim 8
Dependent Claim
22 words
A method according to claim 1, wherein said test sample is contacted with at least one antibody as said specific binding substance.
Claim 9
Dependent Claim
24 words
A method according to claim 7, wherein the said at least one antibody contacted with said test sample is affixed to a solid phase.
Claim 10
Dependent Claim
22 words
A method according to claim 7, wherein the said at least one antibody contacted with said test sample is a monoclonal antibody.
Claim 11
Dependent Claim
60 words
A method according to claim 1, wherein said contacting steps comprises contacting the test sample with a first specific binding substance which binds specifically to said at least one characteristic determinant of said analyte, and with second specific binding substance which binds specifically to said first specific binding substance, said second specific binding substance being affixed to a solid phase.
Claim 12
Dependent Claim
33 words
A method according to claim 10, wherein said test sample is contacted with a first antibody as the first specific binding substance and with a second antibody as the second specific binding substance.
Claim 13
Dependent Claim
32 words
A method according to claim 1, wherein said detectable reporter substance is extractably coupled to said analyte in said coupling step and said reporter substance is extracted therefrom prior to detection thereof.
Claim 14
Dependent Claim
59 words
A method according to claim 1, wherein the detectable reporter substance coupled to said analyte is selected from the group consisting of molecules or ions directly or indirectly detectable based on light absorbance, fluorescence, phosphorescence, or luminescence properties; molecules or ions detectable by their radioactive properties; and molecules or ions detectable by their nuclear magnetic resonance or paramagnetic properties.
Claim 15
Independent Claim
193 words
A method for analyzing a population of cells with lipid-containing membranes to determine the presence or quantity of a subpopulation of said cells having at least one characteristic antigen, within said population, comprising: (i) coupling said cells of said population nonselectively to an extractable reporter substance, said extractable reporter substance having a lipophilic moiety, whereby said lipophilic moiety of said extractable reporter substance stably associates with said lipid-containing membranes of said cells; (ii) contacting said cell population with an antibody which binds specifically to said at least one characteristic antigen of said subpopulation of cells, under conditions causing binding of said antibody to said antigen, said antibody being affixed to magnetic particles; (iii) magnetically separating said magnetic particles bearing said subpopulation of cells from said population; (iv) extracting said reporter substance from said separated subpopulation of cells or from the population remaining after separation of said subpopulation; and (v) detecting the level of extracted reporter substance in said separated subpopulation of cells, or in the population remaining after separation of said subpopulation to provide an indication of the presence or quantity of the subpopulation of cells having said characteristic antigen within said population.
Claim 16
Independent Claim
193 words
A method for analyzing a population of cells with lipid-containing membranes to determine the presence or quantity of a subpopulation of said cells having at least one characteristic antigen, within said population, comprising: (i) coupling said cells of said population nonselectively to an extractable reporter substance, said extractable reporter substance having a lipophilic moiety, whereby said lipophilic moiety of said extractable reporter substance stably associates with said lipid-containing membranes of said cells; (ii) contacting said cell population with an antibody which binds specifically to said at least one characteristic antigen of said subpopulation of cells, under conditions causing binding of said antibody to said antigen, said antibody being affixed to magnetic particles; (iii) magnetically separating said magnetic particles bearing said subpopulation of cells from said population; (iv) extracting said reporter substance from said separated subpopulation of cells or from the population remaining after separation of said subpopulation; and (v) detecting the level of extracted reporter substance in said separated subpopulation of cells, or in the population remaining after separation of said subpopulation to provide an indication of the presence or quantity of the subpopulation of cells having said characteristic antigen within said population.
Claim 17
Dependent Claim
35 words
A method according to claim 14, which also includes the step of relating the level of detected reporter substance to a pre-determined standard to determine the presence or quantity of said subpopulation within said population.
Claim 18
Dependent Claim
43 words
A method according to claim 15, wherein the level of detected reporter substance is related to a standard containing a predetermined quantity of said reporter substance, thereby to determine the number of cells having said at least one characteristic antigen within said population.
Claim 19
Dependent Claim
58 words
A method according to claim 14, wherein the extractable substance coupled to said cells is selected from the group consisting of molecules or ions directly or indirectly detectable based on light absorbance, fluorescence, phosphorescence, or luminescence properties; molecules or ions detectable by their radioactive properties; and molecules or ions detectable by their nuclear magnetic resonance or paramagnetic properties.
Claim 20
Dependent Claim
19 words
A method according to claim 14, wherein said cells are coupled to a fluorochrome as the extractable reporter substance.
Claim 21
Dependent Claim
22 words
A method according to claim 14, wherein said population comprises cells selected from the group consisting of leukocytes, erythrocytes and mixtures thereof.
Claim 22
Dependent Claim
42 words
A method according to claim 14, wherein said population comprises hemopoietic cells and said subpopulation of cells is selected from the group consisting of hemopoietic cells of pre-determined lineage, homopoietic cells of pre-determined function or homopoietic cells of pre-determined stage of differentiation.
Claim 23
Independent Claim
199 words
A method for analyzing a population of cells with lipid-containing membranes to determine the presence or quantity of a subpopulation of said cells having at least one characteristic antigen, within said population, comprising: (i) coupling said cells of said population non-selectively to an extractable reporter substance said extractable reporter substance having a lipophilic moiety, whereby said lipophilic moiety of said extractable reporter substance stably associates with said lipid-containing membranes of said cells; (ii) contacting said cell population with (a) an antibody which binds specifically to said at least one characteristic antigen of said subpopulation of cells and (b) a specific binding substance which binds specifically with said antibody, said specific binding substance being affixed to a solid phase; (iii) separating from said population the solid phase bearing said subpopulation of cells; (iv) extracting said extractable reporter substance from said separated subpopulation of cells or from the population remaining after separation of said subpopulation; and (v) detecting the level of extracted reporter substance in said separated subpopulation of cells, or in the population remaining after separation of said subpopulation to provide an indication of the presence or quantity of the subpopulation of cells having said characteristic antigen within said population.
Claim 24
Independent Claim
199 words
A method for analyzing a population of cells with lipid-containing membranes to determine the presence or quantity of a subpopulation of said cells having at least one characteristic antigen, within said population, comprising: (i) coupling said cells of said population non-selectively to an extractable reporter substance said extractable reporter substance having a lipophilic moiety, whereby said lipophilic moiety of said extractable reporter substance stably associates with said lipid-containing membranes of said cells; (ii) contacting said cell population with (a) an antibody which binds specifically to said at least one characteristic antigen of said subpopulation of cells and (b) a specific binding substance which binds specifically with said antibody, said specific binding substance being affixed to a solid phase; (iii) separating from said population the solid phase bearing said subpopulation of cells; (iv) extracting said extractable reporter substance from said separated subpopulation of cells or from the population remaining after separation of said subpopulation; and (v) detecting the level of extracted reporter substance in said separated subpopulation of cells, or in the population remaining after separation of said subpopulation to provide an indication of the presence or quantity of the subpopulation of cells having said characteristic antigen within said population.
Claim 25
Dependent Claim
35 words
A method according to claim 21, which also includes the step of relating the level of detected reporter substance to a pre-determined standard to determine the presence or quantity of said subpopulation within said population.
Claim 26
Dependent Claim
59 words
A method according to claim 21, wherein the extractable reporter substance coupled to said cells is selected from the group consisting of molecules or ions directly or indirectly detectable based on light absorbance, fluorescence, phosphorescence, or luminescence properties; molecules or ions detectable by their radioactive properties; and molecules or ions detectable by their nuclear magnetic resonance or paramagnetic properties.
Claim 27
Dependent Claim
19 words
A method according to claim 21, wherein said cells are coupled to a fluorochrome as the extractable reporter substance.
Claim 28
Independent Claim
329 words
A method for analyzing a subpopulation of cells, having at least one characteristic determinant, present within a population of cells with lipid-containing membranes, said subpopulation of cells including individual subsets of interest, each said subset having at least one characteristic determinant, to determine the proportion of at least one subset of said cell subpopulation, comprising: (i) non-selectively coupling substantially all of said cells of said population to a detectable reporter substance said detectable reporter substance having a lipophilic moiety, whereby said detectable reporter substance stably associates with said lipid-containing membranes of said cells; (ii) contacting a first sample of said cell population from step (i) with a firs reagent comprising at least one specific binding substance which binds specifically to said at least one characteristic determinant of said cell subpopulation, under conditions causing binding of said first reagent to said cells of said subpopulation, thereby forming first complexes in said first sample; (iii) separating said first complexes in said first sample from unbound cells in said first samples; (iv) detecting the occurrence of said reporter substance in said first complexes; (v) contacting a second sample of said cell population from step (i), of equivalent volume and cell concentration to said first sample, with a second reagent comprising a specific binding substance capable of selectively interacting with said at least one characteristic determinant of said at least one subset of interest in said subpopulation, under conditions causing binding of said second reagent to said at least one characteristic determinant, thereby forming second complexes in said second sample; (vi) separating said second complexes in said second sample from unbound cells in said second sample; (vii) detecting the occurrence of a reporter substance in said second complexes; and (viii) determining the proportion of said at least one subset of interest in said cell subpopulation by quantitating the amount of said reporter substance stably associated with said second complexes relative to the amount of said reporter substance stably associated with said first complexes.
Claim 29
Independent Claim
329 words
A method for analyzing a subpopulation of cells, having at least one characteristic determinant, present within a population of cells with lipid-containing membranes, said subpopulation of cells including individual subsets of interest, each said subset having at least one characteristic determinant, to determine the proportion of at least one subset of said cell subpopulation, comprising: (i) non-selectively coupling substantially all of said cells of said population to a detectable reporter substance said detectable reporter substance having a lipophilic moiety, whereby said detectable reporter substance stably associates with said lipid-containing membranes of said cells; (ii) contacting a first sample of said cell population from step (i) with a firs reagent comprising at least one specific binding substance which binds specifically to said at least one characteristic determinant of said cell subpopulation, under conditions causing binding of said first reagent to said cells of said subpopulation, thereby forming first complexes in said first sample; (iii) separating said first complexes in said first sample from unbound cells in said first samples; (iv) detecting the occurrence of said reporter substance in said first complexes; (v) contacting a second sample of said cell population from step (i), of equivalent volume and cell concentration to said first sample, with a second reagent comprising a specific binding substance capable of selectively interacting with said at least one characteristic determinant of said at least one subset of interest in said subpopulation, under conditions causing binding of said second reagent to said at least one characteristic determinant, thereby forming second complexes in said second sample; (vi) separating said second complexes in said second sample from unbound cells in said second sample; (vii) detecting the occurrence of a reporter substance in said second complexes; and (viii) determining the proportion of said at least one subset of interest in said cell subpopulation by quantitating the amount of said reporter substance stably associated with said second complexes relative to the amount of said reporter substance stably associated with said first complexes.
Claim 30
Dependent Claim
163 words
A method according to claim 25, which includes: (ix) contacting one or more additional samples of said cell population from step (i), each sample being of equivalent volume and cell concentration to said first sample, with one or more additional reagent, each said additional reagent comprising a specific binding substance which binds specifically to at least one characteristic determinant of one or more additional subset of interest respectively, under conditions causing binding of each additional reagent to each said subset, thereby forming additional complexes in said additional samples; (x) separating each said additional complexes in said each additional sample from unbound cells in each said additional sample; (xi) detecting the occurrence of said reporter substance in each said additional complexes; and (xii) determining the proportion of each said additional subsets in said cell subpopulation by quantitating the amount of said reporter substance stably associated with each said additional complex relative to the amount of said reporter substance stably associated with said first complexes.
Claim 31
Dependent Claim
50 words
A method according to claim 25, wherein said first reagent comprises a mixture of specific binding substances, each said binding substance being capable of specifically binding to characteristic determinants of individual subsets of interest comprising said subpopulation, wehreby substantially all cells of said subpopulation are bound to said binding substances.
Claim 32
Dependent Claim
37 words
A method according to claim 25 or 26, which also includes the step of relating the level of detected reported substance to a predetermined standard to determine the presence or quantity of said subset within said subpopulation.
Claim 33
Dependent Claim
31 words
A method according to claim 26, wherein said specific binding substances of said first reagent, said second reagent and said one or more additional reagent being affixed to a solid phase.
Claim 34
Dependent Claim
52 words
A method according to claim 26, wherein said specific binding substances of said first reagent, said second reagent and said one or more additional reagent being affixed to magnetic particles, and said first complexes, said second complexes and said additional complexes bearing said subpopulations of interest are magnetically separated from said samples.
Claim 35
Dependent Claim
30 words
A method according to claim 26, wherein said specific binding substances of said first reagent, said second reagent and said one or more additional reagent comprise at least one antibody.
Claim 36
Dependent Claim
31 words
A method according to claim 26, wherein said specific binding substances of said first reagent, said second reagent and said one or more additional reagent comprise at least one monoclonal antibody.
Claim 37
Dependent Claim
32 words
A method according to claim 25 or 26, wherein said detectable reporter substance is extractably coupled to said cells and said reporter substance is extracted from said cells prior to detection thereof.
Claim 38
Dependent Claim
60 words
A method according to claim 25 or 26, wherein the reporter substance coupled to said cells is selected from the group consisting of molecules or ions directly or indirectly detectable based on light absorbance, fluorescence, phosphorescence, or luminescence properties; molecules or ions detectable by their radioactive properties; and molecules or ions detectable by their nuclear magnetic resonance or paramagnetic properties.
Claim 39
Dependent Claim
20 words
A method according to claim 25 or 26, wherein said detectable reporter substance coupled to said cells comprises a fluorochrome.
Claim 40
Dependent Claim
42 words
A method according to claim 25 which also includes before said separating step (iii), contacting said first sample with an auxiliary specific binding substance which binds specifically to said first reagent, said auxiliary specific binding substance being affixed to a solid phase.
Claim 41
Dependent Claim
42 words
A method according to claim 25, which also includes before said separating step (iii) contacting said second sample with an auxiliary specific binding substance which binds specifically to said second reagent, said auxiliary specific binding substance being affixed to a solid phase.
Claim 42
Dependent Claim
53 words
A method according to claim 26, which also includes before said separating step (vi) contacting at least one of said additional samples with an auxiliary specific binding substance which binds specifically to the additional reagent with which said additional sample is contacted, said auxiliary specific binding substance being affixed to a solid phase.
Claim 43
Dependent Claim
34 words
A method according to claim 36, 37 or 38, wherein said first sample, said second sample or said at least one additional sample are contacted with an antibody as the auxiliary specific binding substance.
Claim 44
Dependent Claim
28 words
A method according to claim 39, wherein the antibody contacted with said first sample, said second sample and said at least one additional sample is a monoclonal antibody.
Claim 45
Independent Claim
70 words
A method for analyzing a population of cells with lipid-containing membranes to determine the presence or quantity of a subpopulation of said cells, having a characteristic determinant, within said population, comprising: (i) coupling said cells of said population non-selectively to an extractable reporter substance, said extractable reporter substance having a lipophilic moiety, whereby said lipophilic moiety of said extractable reporter substance stably associates with said lipid-containing membranes of said cells;
Claim 46
Independent Claim
70 words
A method for analyzing a population of cells with lipid-containing membranes to determine the presence or quantity of a subpopulation of said cells, having a characteristic determinant, within said population, comprising: (i) coupling said cells of said population non-selectively to an extractable reporter substance, said extractable reporter substance having a lipophilic moiety, whereby said lipophilic moiety of said extractable reporter substance stably associates with said lipid-containing membranes of said cells;
Claim 47
Dependent Claim
90 words
(ii) contacting said cell population with a specific binding substance which binds specifically to said determinant under conditions causing binding of said specific binding substance to said determinant, said specific binding substance being affixed to a solid phase; (iii) separating said solid phase, bearing said subpopulation, from said population; (iv) extracting said reporter substance from said separated subpopulation of cells; and (v) detecting the level of extracted reporter substance to provide an indication of the presence or quantity of the subpopulation of cells having said characteristic determinant within said population.
Claim 48
Dependent Claim
90 words
(ii) contacting said cell population with a specific binding substance which binds specifically to said determinant under conditions causing binding of said specific binding substance to said determinant, said specific binding substance being affixed to a solid phase; (iii) separating said solid phase, bearing said subpopulation, from said population; (iv) extracting said reporter substance from said separated subpopulation of cells; and (v) detecting the level of extracted reporter substance to provide an indication of the presence or quantity of the subpopulation of cells having said characteristic determinant within said population.
Claim 49
Dependent Claim
35 words
A method according to claim 41, which also includes the step of relating the level of detected reporter substance to a pre-determined standard to determine the presence or quantity of said subpopulation within said population.
Claim 50
Dependent Claim
58 words
A method according to claim 41, wherein the reporter substance coupled to said analyte is selected from the group consisting of molecules or ions directly or indirectly detectable based on light absorbance, fluorescence, phosphorescence, or luminescence properties; molecules or ions detectable by their radioactive properties; and molecules or ions detectable by their nuclear magnetic resonance or paramagnetic properties.
Claim 51
Dependent Claim
19 words
A method according to claim 41, wherein said cells are coupled to an extractable fluorochrome as the reporter substance.
Claim 52
Dependent Claim
38 words
A method according to claim 41, wherein said subpopulation has at least one antigen as said at least one characteristic determinant and said cell population is contacted with an antibody to said antigen, as the specific binding substance.
Claim 53
Dependent Claim
18 words
A method according to claim 45, wherein the antibody contacted with said cell population is a monoclonal antibody.
Claim 54
Independent Claim
44 words
A method for determining the presence or quantity of a subpopulation of biological entities selected from the group consisting of eucaryotic cells, procaryotic cells and viruses, and having at least one characteristic determinant, within a population of said entities in a test sample, comprising:
Claim 55
Independent Claim
44 words
A method for determining the presence or quantity of a subpopulation of biological entities selected from the group consisting of eucaryotic cells, procaryotic cells and viruses, and having at least one characteristic determinant, within a population of said entities in a test sample, comprising:
Claim 56
Dependent Claim
74 words
(i) coupling a detectable reporter substance non-selectively to said population of biological entities; (ii) contacting said biological entities in said test sample with a specific binding substance which binds specifically to said at least one characteristic determinant of said subpopulation of biological entities, under conditions causing binding of said binding substance to said at least one characteristic determinant; (iii) separating said specific binding substance bearing said subpopulation of biological entities, from said population; and
Claim 57
Dependent Claim
74 words
(i) coupling a detectable reporter substance non-selectively to said population of biological entities; (ii) contacting said biological entities in said test sample with a specific binding substance which binds specifically to said at least one characteristic determinant of said subpopulation of biological entities, under conditions causing binding of said binding substance to said at least one characteristic determinant; (iii) separating said specific binding substance bearing said subpopulation of biological entities, from said population; and
Claim 58
Dependent Claim
92 words
(iv) simultaneously detecting the occurrence of said detectable reporter substance in a plurality of said biological entities in said separated subpopulation of biological entities in said test sample or in a plurality of said biological entities in said population of said entities remaining in said test sample after separation of said subpopulation, thereby to provide an indication determinative of the presence or quantity of said subpopulation of biological entities having said at least one characteristic determinant within said population of biological entities having said at least one characteristic determinant within said population.
Claim 59
Dependent Claim
92 words
(iv) simultaneously detecting the occurrence of said detectable reporter substance in a plurality of said biological entities in said separated subpopulation of biological entities in said test sample or in a plurality of said biological entities in said population of said entities remaining in said test sample after separation of said subpopulation, thereby to provide an indication determinative of the presence or quantity of said subpopulation of biological entities having said at least one characteristic determinant within said population of biological entities having said at least one characteristic determinant within said population.
Claim 60
Dependent Claim
35 words
A method according to claim 47, which also includes the step of relating the level of detected reporter substance to a predetermined standard to determine the presence or quantity of said subpopulation within said population.
Claim 61
Dependent Claim
42 words
A method according to claim 47, wherein the level of detected reporter substance is related to a standard containing a predetermined quantity of said reporter substance, thereby to determine the number of said biological entities having said characteristic determinant within said population.
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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) | 1996-10-25 | 1997-04-26 | 1997-10-26 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2000-10-25 | 2001-04-26 | 2001-10-26 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2004-10-25 | 2005-04-26 | 2005-10-26 | $7700.00 | Window Closed / Lapsed |