EP0143574A1
Expired / Lapsed
EP
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Assay method and kit for whole blood samples
Assignee: Siemens Healthcare Diagnostics GmbH Germany
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Filing: 1984-11-01
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Granted: —
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Expires: 2004-11-01
Abstract
Abstract @ Methods and assay kits are provided for detecting the presence of a member of a specific binding pair (sbp) in a whole blood sample. The sbp members consist of ligand and homologous receptor. In the method of the invention the whole blood sample, without separation of red blood cells, is combined in an aqueous buffered medium with a binding agent for the red blood cells in the sample. Where appropriate to the assay protocol, none or one or more members of a signal producing system, which is capable of producing a detectible signal, are included. At least one of the sbp members is substantially uniformly bound to a solid bibulous element. When the assay medium is contacted with a predetermined region of the solid bibulous element, the red blood cells will concentrate in an area on the bibulous element to the air/liquid interface. Furthermore, the medium will traverse the solid bibulous element. The remaining members of the signal producing system are then combined with the solid bibulous element. The signal produced as a result of the binding of the sbp members is detected and is related to the amount of the sbp member in the sample.
Inventors
- 👤 Robert F. Zuk
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-31 08:09
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/80; G01N33/53; G01N33/543; G01N33/54366; G01N33/54386; G01N33/54387; G01N33/54388; Y; Y10; Y10S; Y10S435/00; Y10S435/805; Y10S435/962; Y10S435/966; Y10S435/97; Y10S435/975; Y10S436/00; Y10S436/807; Y10S436/808; Y10S436/81; Y10S436/825; Y10S436/827
Claims Analysis
Total Claims: 14
Claim 1
Independent Claim
131 words
A method for determining an analyte in a whole blood sample, said analyte being a member of a specific binding pair ("sbp member") consisting of ligand and homologous receptor, which method comprises - contacting in an aqueous medium (A) a bibulous element, at a predetermined region of said element defining an air-liquid interface, said element having an sbp member substantially uniformly bound to the surface thereof with (B) said whole blood sample and (C) a binding agent for red blood cells present in said whole blood sample whereby said medium traverses said element to define a border related to the amount of analyte and said red blood cells aggregate at said air-liquid interface; and determining said border by means of a signal producing system, which system includes a labeled sbp member.
Claim 2
Independent Claim
131 words
A method for determining an analyte in a whole blood sample, said analyte being a member of a specific binding pair ("sbp member") consisting of ligand and homologous receptor, which method comprises - contacting in an aqueous medium (A) a bibulous element, at a predetermined region of said element defining an air-liquid interface, said element having an sbp member substantially uniformly bound to the surface thereof with (B) said whole blood sample and (C) a binding agent for red blood cells present in said whole blood sample whereby said medium traverses said element to define a border related to the amount of analyte and said red blood cells aggregate at said air-liquid interface; and determining said border by means of a signal producing system, which system includes a labeled sbp member.
Claim 3
Dependent Claim
13 words
The method of Claim 1 wherein the binding agent is an agglutinating agent.
Claim 4
Dependent Claim
15 words
The method of Claim 1 wherein the binding agent is specific for red blood cells.
Claim 5
Dependent Claim
16 words
The method of Claim 1 wherein the binding agent is antibody specific for red blood cells.
Claim 6
Dependent Claim
12 words
The method of Claim 1 wherein the binding agent is a lectin.
Claim 7
Dependent Claim
14 words
The method of Claim 1 wherein the binding agent is a polymeric amino acid.
Claim 8
Dependent Claim
11 words
The method of Claim 1 wherein the binding agent is polylysine.
Claim 9
Dependent Claim
15 words
The method of Claim 1 wherein the pH is in the range of about 6-10.
Claim 10
Dependent Claim
19 words
The method of Claim 1 wherein the ionic strength in Step b is in the range of about 0.1-0.4.
Claim 11
Independent Claim
196 words
A method for detecting the presence of_ a member of a specific binding pair ("sbp member") in a whole blood sample, said sbp member consisting of ligand and homologous receptor, where said method involves (1) a binding agent for red blood cells in said sample; (2) a solid bibulous element to which is bound at least one sbp member; and (3) a signal producing system capable of producing a detectable signal, the members of which include at least one label which is bound to an sbp member, said method comprising: (a) combining in an aqueous medium said sample, said binding agent, and fewer than all the members of the signal producing system, (b) contacting a portion of said bibulous carrier with said aqueous medium to produce an air-liquid interface and to allow said medium to diffuse along said bibulous element and said red blood cells to concentrate in an area adjacent the air/liquid interface, (c) contacting said bibulous element with any remaining members of the signal producing system, and (d) detecting the signal as a result of said signal producing system, wherein said signal is related to the amount of said sbp member in said sample.
Claim 12
Independent Claim
196 words
A method for detecting the presence of_ a member of a specific binding pair ("sbp member") in a whole blood sample, said sbp member consisting of ligand and homologous receptor, where said method involves (1) a binding agent for red blood cells in said sample; (2) a solid bibulous element to which is bound at least one sbp member; and (3) a signal producing system capable of producing a detectable signal, the members of which include at least one label which is bound to an sbp member, said method comprising: (a) combining in an aqueous medium said sample, said binding agent, and fewer than all the members of the signal producing system, (b) contacting a portion of said bibulous carrier with said aqueous medium to produce an air-liquid interface and to allow said medium to diffuse along said bibulous element and said red blood cells to concentrate in an area adjacent the air/liquid interface, (c) contacting said bibulous element with any remaining members of the signal producing system, and (d) detecting the signal as a result of said signal producing system, wherein said signal is related to the amount of said sbp member in said sample.
Claim 13
Dependent Claim
17 words
The method of Claim 10 wherein the binding agent is antibody specific for said red blood cells.
Claim 14
Dependent Claim
77 words
A kit for use in the method of Claim 1, said kit comprising in a packaged combination in predetermined ratios for combination with said sample according to said method, (a) a binding agent for said red blood cells, (b) a bibulous element to which is bound at least one sbp member, (c) a signal producing system capable of producing a detectable signal in relation to the amount of analyte present in said sample, and (d) ancillary materials.
References Cited (Backward Citations)
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