Patent EP0176252A2 successfully refreshed from Google Patents.
EP0176252A2
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Assay method employing fluorescent cell incorporative dye
Assignee: Syntex USA LLC
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Filing: 1985-08-27
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Granted: —
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Expires: 2005-08-27
Abstract
Abstract A method is provided for determining the presence in a sample of a member of a specific binding pair ("sbp member") consisting of ligand and its homologous receptor. The sample is combined in an aqueous medium with (1) a complementary sbp member wherein at least the sbp member or the complementary sbp member is bound to the surface of a cell and (2) a fluorescent agent capable of being incorporated into the cell. The presence of the sbp member is indicated by a change in fluorescence of the unseparated cell suspension as a result of agglutination of the cells. The present invention has particular application to blood typing, for example, for the determination of the presence of blood group antigens A, B, AB, 0, and D (Rh o ) and antibodies to such antigens.
Inventors
- 👤 Chiu C. Chang
- 👤 Vartan Ghazarossian
- 👤 Edwin F. Ullman
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-07 15:49
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/543; G01N33/554; G01N33/555; G01N33/531; G01N33/532; G01N33/533; G01N33/536; G01N33/542; G01N33/80; Y; Y10; Y10S; Y10S436/00; Y10S436/80; Y10S436/805; Y10S436/807; Y10S436/829
Claims Analysis
Total Claims: 18
Claim 1
Independent Claim
111 words
A method for determining in a sample the presence of a member of a specific binding pair ("sbp member") consisting of ligand and its homologous receptor, which comprises - (a) combining in an aqueous medium (1) said sample, (2) a complementary sbp member wherein at least one of said sbp member or said complementary sbp member is bound to the surface of a cell and (3) a fluorescent agent capable of being incorporated into said cell and (b) without separating said cells from said aqueous medium determining a change in fluorescence as a result of agglutination of said cells, said change being an indication of the presence of said sbp member.
Claim 2
Independent Claim
111 words
A method for determining in a sample the presence of a member of a specific binding pair ("sbp member") consisting of ligand and its homologous receptor, which comprises - (a) combining in an aqueous medium (1) said sample, (2) a complementary sbp member wherein at least one of said sbp member or said complementary sbp member is bound to the surface of a cell and (3) a fluorescent agent capable of being incorporated into said cell and (b) without separating said cells from said aqueous medium determining a change in fluorescence as a result of agglutination of said cells, said change being an indication of the presence of said sbp member.
Claim 3
Dependent Claim
14 words
The method of claim 1 wherein said sbp member is a cell surface member.
Claim 4
Dependent Claim
20 words
The method of claim 1 or claim 2 wherein said complementary sbp member is a receptor for a surface antigen.
Claim 5
Independent Claim
18 words
The method of any one of claims 1 to 3 wherein said cell is a red blood cell.
Claim 6
Independent Claim
28 words
The method of any one of claims 1 to 4 wherein said change in fluorescence is determined by a non-flow cytometry technique, preferably by a fiber optic cytometer.
Claim 7
Independent Claim
23 words
The method of any one of claims 1 to 5 wherein said fluorescent agent is a membrane soluble dye, preferably a squarate dye.
Claim 8
Independent Claim
97 words
A method for determining in a sample of whole blood the presence of a member of a specific binding pair ("sbp member"), which comprises - (a) combining in an aqueous medium (1) said sample, (2) a complementary sbp member wherein at least one of said sbp member or said complementary sbp member is bound to the surface of a cell and (3) a fluorescent agent capable of being incorporated into said cells and (b) without separating said cells from said aqueous medium, determining a change in fluorescence as an indication of the presence of said sbp member.
Claim 9
Independent Claim
97 words
A method for determining in a sample of whole blood the presence of a member of a specific binding pair ("sbp member"), which comprises - (a) combining in an aqueous medium (1) said sample, (2) a complementary sbp member wherein at least one of said sbp member or said complementary sbp member is bound to the surface of a cell and (3) a fluorescent agent capable of being incorporated into said cells and (b) without separating said cells from said aqueous medium, determining a change in fluorescence as an indication of the presence of said sbp member.
Claim 10
Dependent Claim
23 words
The method of claim 7 wherein said change in fluorescence is determined by a non-flow cytometric technique, preferably by a fiber optic cytometer.
Claim 11
Dependent Claim
21 words
The method of claim 7 or claim 8 wherein said fluorescent agent is a membrane soluble dye, preferably a squarate dye.
Claim 12
Independent Claim
25 words
The method of any one of claims 7 to 9 wherein said cells are red blood cells, and said sbp member is a surface antigen.
Claim 13
Dependent Claim
23 words
The method of claim 10 wherein said surface antigen is selected from the group of antigens consisting of A, B, D and immunoglobulins.
Claim 14
Independent Claim
23 words
The method of any one of claims 7 to 11 wherein said change in fluorescence is a result of agglutination of said cells.
Claim 15
Dependent Claim
198 words
The method of claim 7 wherein said fluorescent agent is a squarate dye of the formula selected from wherein R 1 , R 2 , R 3 , and R 4 are independently selected from alkyl of from 2 to 16 carbon atoms and aralkyl of from 2 to 16 carbon atoms wherein the groups may be the same or different and may be substituted with a substituent selected from OH, CONHR 1 , SR 1 , OR 1 wherein R l is lower alkyl of from 1 to 6 carbon atoms, F, Cl, Br, I, N0 2 , oxocarbonyl and CN, R 5 , and R 6 are independently selected from hydrogen, methoxy, and hydroxyl, or wherein R 1 , R 2 , and R 3 are independently selected from alkyl of from 2 to 16 carbon atoms and aralkyl of from 2 to 16 carbon atoms wherein the groups may be the same or different and may be substituted with a substituent selected from OH, CONHR 1 , SR 1 , OR 1 wherein R l is lower alkyl of from 1 to 6 carbon atoms, F, Cl, Br, I, N0 2 , oxocarbonyl, and CN.
Claim 16
Independent Claim
31 words
A method for typing red blood cells wherein an agglutination of said cells is employed, characterized by combining said cells with a fluorescent agent capable of being incorporated into said cells.
Claim 17
Independent Claim
31 words
A method for typing red blood cells wherein an agglutination of said cells is employed, characterized by combining said cells with a fluorescent agent capable of being incorporated into said cells.
Claim 18
Dependent Claim
31 words
The method of claim 14 wherein said fluorescent agent is a squarate dye having an absorption wavelength of greater than 600 nanometers, preferably a squarate dye as defined in claim 13.
References Cited (Backward Citations)
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