Patent EP0595641A2 successfully refreshed from Google Patents.
EP0595641A2
Expired / Lapsed
EP
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One-step simultaneous immunoassay
Assignee: Wardlaw Stephen Clark
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Filing: 1993-10-29
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Granted: —
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Expires: 2013-10-29
Abstract
Abstract A patient's health is diagnosed by centrifuging blood samples in a transparent tube, which tube contains one or more groups of particles such as lyposomes or plastic beads of different densities for each group. Each group of density-defined particles carries antigens or antibodies which are specific to a complement antigen or antibody which may be in the blood sample being tested, and which are indicative of the patient's health. A label-tagged antibody which is specific to all bound antibody/antigen couples is added to the blood sample so as to form labeled antibody+antigen-antibody complexes (AAAC) in the blood sample. Upon centrifugation, the complexed particles will settle out in different areas in the tube according to the respective density of the particles, and the degree of label emission of the particle layers can enable qualitative or quantitative analyses of the blood sample to be made. Unbound labeled antibodies will be washed away from the complexed layers by the washing action of the descending blood cells during the centrifugation step. Unbound labeled antibodies will thus not interfere with the analysis.
Inventors
- 👤 Robert A. Levine
- 👤 Stephen C. Wardlaw
- 👤 Leon W. M. M. Terstappen
- 👤 Diether J. Recktenwald
- 👤 Thomas Mercolino
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-23 11:29
CPC Classifications
B; B01; B01L; B01L3/00; B01L3/50; B01L3/502; B01L3/5021; G; G01; G01N; G01N33/00; G01N33/48; G01N33/483; G01N33/487; G01N33/49; G01N33/491; G01N33/50; G01N33/53; G01N33/536; G01N33/537; G01N33/5375; G01N33/569; G01N33/56966; G01N33/56972; G01N33/58; G01N33/585; Y; Y02; Y02A; Y02A50/00; Y02A50/30; Y10; Y10S; Y10S435/00; Y10S435/81; Y10S435/967; Y10S435/971; Y10S435/973; Y10S436/00; Y10S436/805; Y10S436/807; Y10S436/81; Y10S436/824; Y10S436/829; Y10T; Y10T436/00; Y10T436/11; Y10T436/111666
Claims Analysis
Total Claims: 15
Claim 1
Independent Claim
120 words
A method for detecting a suspect target analyte in a biological fluid sample in a transparent tube, said method comprising the steps of: a) adding a group of density markers to the sample, which density markers have a predetermined specific gravity, each density marker in said group being coupled with a binding material to form density marker couples which are specific to the suspect target analyte; b) adding labeled couple-binding antibodies to the sample; c) incubating the density marker/labeled couple-binding antibody sample mixture; d) densimetrically aggregating the density markers into at least one distinct band in the tube; and e) determining if the band exhibits the presence of the labeled couple-binding antibodies, and therefore the presence of the target analyte.
Claim 2
Independent Claim
120 words
A method for detecting a suspect target analyte in a biological fluid sample in a transparent tube, said method comprising the steps of: a) adding a group of density markers to the sample, which density markers have a predetermined specific gravity, each density marker in said group being coupled with a binding material to form density marker couples which are specific to the suspect target analyte; b) adding labeled couple-binding antibodies to the sample; c) incubating the density marker/labeled couple-binding antibody sample mixture; d) densimetrically aggregating the density markers into at least one distinct band in the tube; and e) determining if the band exhibits the presence of the labeled couple-binding antibodies, and therefore the presence of the target analyte.
Claim 3
Dependent Claim
114 words
A method of Claim 1 wherein one or more different target analytes may be detected in a biological fluid sample in a transparent tube through the addition of distinct groups of density markers to the sample, there being one group of density markers for each target analyte suspected to be in the sample, each group of density markers having a different specific gravity from each other group of density markers, and each density marker in each group thereof being coupled with a binding material which is specific to one of the target analytes, whereby each of the different density marker/binding material couple groups is specific to a different one of the suspected target analytes.
Claim 4
Dependent Claim
28 words
The method of Claim 1 or Claim 2 further comprising the step of displacing unbound labeled couple-binding antibodies from the density markers during the step of densimetrically separating.
Claim 5
Dependent Claim
34 words
The method of Claim 3 wherein the fluid sample is whole blood, and wherein the density-markers have a specific gravity that is greater than the specific gravity of the lightest of the red cells.
Claim 6
Dependent Claim
22 words
The method of Claim 4 wherein said step of displacing is performed by the centrifuged red cells in the whole blood sample.
Claim 7
Dependent Claim
83 words
Apparatus for use in centrifugally analyzing a biological fluid sample for the presence or absence of a target analyte, said apparatus including: a) a transparent tube having a bore for receiving the fluid sample; b) means for forming a local constriction in said tube bore; c) a quantity of target analyte-specific antibody and/or antigen-coupled density-markers in said tube; and d) means forming a density gradient layer in said tube into which the density markers sink during centrifugation of the sample in said tube.
Claim 8
Dependent Claim
83 words
Apparatus for use in centrifugally analyzing a biological fluid sample for the presence or absence of a target analyte, said apparatus including: a) a transparent tube having a bore for receiving the fluid sample; b) means for forming a local constriction in said tube bore; c) a quantity of target analyte-specific antibody and/or antigen-coupled density-markers in said tube; and d) means forming a density gradient layer in said tube into which the density markers sink during centrifugation of the sample in said tube.
Claim 9
Dependent Claim
39 words
The apparatus of Claim 6 further comprising an operable amount of labeled anti-antigenantibody-complex (AAAC) antibodies in said tube, said AAAC antibodies being specific to any antibody/antigen/analyte couples formed on said density-markers and being operable to detectably highlight said couples.
Claim 10
Dependent Claim
51 words
The apparatus of Claim 7 wherein the antibody and/or antigen-coupled density markers and labeled AAAC antibodies are present in the tube in an amount which exceeds the amount of target analyte units expected to be found in the sample to an extent sufficient to quantify the target analyte in the sample.
Claim 11
Dependent Claim
28 words
The apparatus of Claim 8 wherein the amount of antibody and/or antigen-coupled density-markers and labeled AAAC antibodies is at least twice the expected quantity of the target analyte.
Claim 12
Dependent Claim
112 words
Apparatus for use in centrifugally analyzing a biological fluid sample for the presence or absence of immune globulin subgroups, said apparatus including: a) a transparent tube having a bore for receiving the fluid sample; b) means for forming a local constriction in said tube bore; c) a quantity of immune globulin-specific antibody and/or antigen-coupled density-markers in said tube; d) an operable amount of labeled anti-antigenantibody-complex (AAAC) antibodies in said tube, said AAAC antibodies being specific to immune globulin antibody-antigen complexes formed on said density-markers and being operable to detectably highlight said complexes; and e) means for forming a density gradient layer in said tube into which said density-markers will sink during centrifugation.
Claim 13
Dependent Claim
112 words
Apparatus for use in centrifugally analyzing a biological fluid sample for the presence or absence of immune globulin subgroups, said apparatus including: a) a transparent tube having a bore for receiving the fluid sample; b) means for forming a local constriction in said tube bore; c) a quantity of immune globulin-specific antibody and/or antigen-coupled density-markers in said tube; d) an operable amount of labeled anti-antigenantibody-complex (AAAC) antibodies in said tube, said AAAC antibodies being specific to immune globulin antibody-antigen complexes formed on said density-markers and being operable to detectably highlight said complexes; and e) means for forming a density gradient layer in said tube into which said density-markers will sink during centrifugation.
Claim 14
Independent Claim
92 words
A method for detecting a suspect target analyte in a sample, which comprises contacting the sample with (i) a group of density markers coupled with a binding material which binds specifically to the target analyte; and (ii) labelled binding material which binds specifically to the target analyte; separating the density markers and coupled binding material, and any target analyte and labelled binding material bound thereto from unbound labelled binding material by centrifuging the sample, and determining the presence and/or the amount of target analyte in the sample by detection of the label.
Claim 15
Independent Claim
92 words
A method for detecting a suspect target analyte in a sample, which comprises contacting the sample with (i) a group of density markers coupled with a binding material which binds specifically to the target analyte; and (ii) labelled binding material which binds specifically to the target analyte; separating the density markers and coupled binding material, and any target analyte and labelled binding material bound thereto from unbound labelled binding material by centrifuging the sample, and determining the presence and/or the amount of target analyte in the sample by detection of the label.
References Cited (Backward Citations)
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