US4900686A
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Fluorescent conjugates and biological diagnostic assay
Assignee: Siemens Healthcare Diagnostics GmbH Germany
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Filing: 1987-04-02
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Granted: 1990-02-13
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Expires: 2007-04-02
Abstract
Abstract There are provided novel fluorescent conjugates comprising a rhodamine dye moiety and a biologically active moiety. In addition, there are provided methods for determining an analyte in a biological fluid.
Inventors
- 👤 Michael J. Arnost
- 👤 Frank A. Meneghini
- 👤 Paul S. Palumbo
- 👤 Stephen G. Stroud
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-30 22:24
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/531; G01N33/532; G01N33/533; C; C09; C09B; C09B11/00; C09B11/04; C09B11/10; C09B11/24; Y; Y10; Y10S; Y10S435/00; Y10S435/805; Y10S435/81; Y10S436/00; Y10S436/80; Y10S436/805; Y10S436/807; Y10S436/808
Claims Analysis
Total Claims: 33
Claim 1
Independent Claim
282 words
A fluorescent diagnostic assay comprising combining in an assay medium: a sample suspected of containing an analyte; a binding partner for said analyte; and a conjugate of said analyte or an analogue thereof and a fluorescent compound, said conjugate represented by the formula ##STR27## wherein W, X, Y and Z each independently is --(--CH2) a (CHR 4 )(CH 2 --)-- b ; R, R 1 , R 3 and R 4 each independently is hydrogen, a hydrophilic group or a substituent including said analyte or an analogue thereof; R 2 is --CO 2 .sup.⊖, SO 3 .sup.⊖ or a substituent including said analyte or an analogue thereof; provided that one of R, R 1 , R 2 , R 3 and R 4 is a substituent including said analyte or an analogue thereof and at least one of the remaining of R, R 1 , R 3 and R 4 is a hydrophilic group; a and b each independently is an integer of from 0 to 4, provided that the sum of a+b is an integer of from 1 to 4; m and n each is 0 to 1 provided that one of m and n is 0 and the other is 1; p and q each is 0 or 1 provided that one of p and q is 0 and the other is 1; A is a counterion or the counterions to balance the overall charges on the conjugate moiety; and x is 0 or 1; separating bound conjugate from free conjugate; measuring the level of the fluorescent signal of said bound or free conjugate; and relating said level of said signal to the amount of said analyte in said sample.
Claim 2
Dependent Claim
282 words
A fluorescent diagnostic assay comprising combining in an assay medium: a sample suspected of containing an analyte; a binding partner for said analyte; and a conjugate of said analyte or an analogue thereof and a fluorescent compound, said conjugate represented by the formula ##STR27## wherein W, X, Y and Z each independently is --(--CH2) a (CHR 4 )(CH 2 --)-- b ; R, R 1 , R 3 and R 4 each independently is hydrogen, a hydrophilic group or a substituent including said analyte or an analogue thereof; R 2 is --CO 2 .sup.⊖, SO 3 .sup.⊖ or a substituent including said analyte or an analogue thereof; provided that one of R, R 1 , R 2 , R 3 and R 4 is a substituent including said analyte or an analogue thereof and at least one of the remaining of R, R 1 , R 3 and R 4 is a hydrophilic group; a and b each independently is an integer of from 0 to 4, provided that the sum of a+b is an integer of from 1 to 4; m and n each is 0 to 1 provided that one of m and n is 0 and the other is 1; p and q each is 0 or 1 provided that one of p and q is 0 and the other is 1; A is a counterion or the counterions to balance the overall charges on the conjugate moiety; and x is 0 or 1; separating bound conjugate from free conjugate; measuring the level of the fluorescent signal of said bound or free conjugate; and relating said level of said signal to the amount of said analyte in said sample.
Claim 3
Dependent Claim
102 words
The assay as defined in claim 1 wherein said conjugate is represented by the formula ##STR28## where R 12 , R 13 , R 14 and R 15 each independently is hydrogen, a hydrophilic group or a substituent including said analyte or an analogue thereof, provided that one of R 2 , R 3 , R 12 , R 13 , R 14 and R 15 is a substituent including said analyte or an analogue thereof and at least one of the remaining of R 3 , R 12 , R 13 , R 14 and R 15 is a hydrophilic group.
Claim 4
Dependent Claim
20 words
The assay as defined in claim 2 wherein R 2 is a substituent including said analyte or an analogue thereof.
Claim 5
Dependent Claim
14 words
The assay as defined in claim 3 wherein R 12 is a hydrophilic group.
Claim 6
Dependent Claim
14 words
The assay as defined in claim 4 wherein R 13 is a hydrophilic group.
Claim 7
Dependent Claim
49 words
The assay as defined in any of claims 4 and 5 wherein said hydrophilic group is a carboxylic acid, a polyether, a polyalcohol, an amine, a sulfonic acid, a phosphonic acid, a phosphonic ester, a phosphate, a phosphate ester, a phosphinic acid, a boronic acid or a borinic acid.
Claim 8
Dependent Claim
102 words
The assay as defined in claim 1 wherein said conjugate is represented by the formula ##STR29## wherein R 12 , R 13 , R 14 and R 15 each independently is hydrogen, a hydrophilic group or a substituent including said analyte or an analogue thereof, provided that one of R 2 , R 3 , R 12 , R 13 , R 14 and R 15 is a substituent including said analyte or an analogue thereof and at least one of the remaining of R 3 , R 12 , R 13 , R 14 and R 15 is a hydrophilic group.
Claim 9
Dependent Claim
20 words
The assay as defined in claim 7 wherein R 2 is a substituent including said analyte or an analogue thereof.
Claim 10
Dependent Claim
14 words
The assay as defined in claim 8 wherein R 12 is a hydrophilic group.
Claim 11
Dependent Claim
14 words
The assay as defined in claim 9 wherein R 13 is a hydrophilic group.
Claim 12
Dependent Claim
49 words
The assay as defined in any of claims 9 and 10 wherein said hydrophilic group is a carboxylic acid, a polyether, a polyalcohol, an amine, a sulfonic acid, a phosphonic acid, a phosphonic ester, a phosphate, a phosphate ester, a phosphinic acid, a boronic acid or a borinic acid.
Claim 13
Dependent Claim
280 words
A diagnostic assay element adapted to receive a sample of a biological fluid and to provide a detectable signal as a function of an analyte which may be present in said fluid, said assay element including a fluorescent conjugate represented by the formula ##STR30## wherein W, X, Y and Z each independently is --(--CH2) a (CHR 4 )(CH 2 --)-- b ; R, R 1 , R 3 and R 4 each independently is hydrogen, a hydrophilic group or a substituent including a biologically active moiety selected from the group consisting of an antigen, an antibody, a hapten, an Fab fragment and a DNA probe, wherein said biologically active moiety is an analyte of interest, an analogue thereof or a binding partner of said analyte; R 2 is --CO 2 .sup.⊖, SO 3 .sup.⊖ or a substituent including said biologically active moiety; provided that one of R, R 1 , R 2 , R 3 and R 4 is a substituent including said biologically active moiety and at least one of the remaining of R, R 1 , R 3 and R 4 is a hydrophilic group; a and b each independently is an integer of from 0 to 4, provided that the sum of a+b is an integer of from 1 to 4; m and n each is 0 to 1 provided that one of m and n is 0 and the other is 1; p and q each is 0 or 1 provided that one of p and q is 0 and the other is 1; A is a counterion or the counterions to balance the overall charges on the conjugate moiety; and x is 0 or 1.
Claim 14
Dependent Claim
280 words
A diagnostic assay element adapted to receive a sample of a biological fluid and to provide a detectable signal as a function of an analyte which may be present in said fluid, said assay element including a fluorescent conjugate represented by the formula ##STR30## wherein W, X, Y and Z each independently is --(--CH2) a (CHR 4 )(CH 2 --)-- b ; R, R 1 , R 3 and R 4 each independently is hydrogen, a hydrophilic group or a substituent including a biologically active moiety selected from the group consisting of an antigen, an antibody, a hapten, an Fab fragment and a DNA probe, wherein said biologically active moiety is an analyte of interest, an analogue thereof or a binding partner of said analyte; R 2 is --CO 2 .sup.⊖, SO 3 .sup.⊖ or a substituent including said biologically active moiety; provided that one of R, R 1 , R 2 , R 3 and R 4 is a substituent including said biologically active moiety and at least one of the remaining of R, R 1 , R 3 and R 4 is a hydrophilic group; a and b each independently is an integer of from 0 to 4, provided that the sum of a+b is an integer of from 1 to 4; m and n each is 0 to 1 provided that one of m and n is 0 and the other is 1; p and q each is 0 or 1 provided that one of p and q is 0 and the other is 1; A is a counterion or the counterions to balance the overall charges on the conjugate moiety; and x is 0 or 1.
Claim 15
Dependent Claim
18 words
The diagnostic element as defined in claim 12 which includes a support carrying at least one reagent layer.
Claim 16
Dependent Claim
22 words
The diagnostic element as defined in claim 12 which includes the binding partner of said biologically active moiety present in said conjugate.
Claim 17
Dependent Claim
99 words
The assay element as defined in claim 12 wherein said conjugate is represented by the formula ##STR31## wherein R 12 , R 13 , R 14 and R 15 each independently is hydrogen, a hydrophilic group or a substituent including said biologically active moiety, provided that one of R 2 , R 3 , R 12 , R 13 , R 14 and R 15 is a substituent including said biologically active moiety and at least one of the remaining of R 3 , R 12 , R 13 , R 14 and R 15 is a hydrophilic group.
Claim 18
Dependent Claim
19 words
The assay element as defined in claim 15 wherein R 2 is a substituent including said biologically active moiety.
Claim 19
Dependent Claim
15 words
The assay element as defined in claim 16 wherein R 12 is a hydrophilic group.
Claim 20
Dependent Claim
15 words
The assay element as defined in claim 17 wherein R 13 is a hydrophilic group.
Claim 21
Dependent Claim
50 words
The assay element as defined in any of claims 17 or 18 wherin said hydrophilic group is a carboxylic acid, a polyether, a polyalcohol, an amine, a sulfonic acid, a phosphonic acid, a phosphonic ester, a phosphate, a phosphate ester, a phosphinic acid, a boronic acid or a borinic acid.
Claim 22
Dependent Claim
99 words
The assay element as defined in claim 12 wherein said conjugate is represented by the formula ##STR32## wherein R 12 , R 13 , R 14 and R 15 each independently is hydrogen, a hydrophilic group or a substituent including said biologically active moiety, provided that one of R 2 , R 3 , R 12 , R 13 , R 14 and R 15 is a substituent including said biologically active moiety and at least one of the remaining of R 3 , R 12 , R 13 , R 14 and R 15 is a hydrophilic group.
Claim 23
Dependent Claim
19 words
The assay element as defined in claim 20 wherein R 2 is a substituent including said biologically active moiety.
Claim 24
Dependent Claim
15 words
The assay element as defined in claim 21 wherein R 12 is a hydrophilic group.
Claim 25
Dependent Claim
15 words
The assay element as defined in claim 22 wherein R 13 is a hydrophilic group.
Claim 26
Dependent Claim
50 words
The assay element as defined in any of claims 22 or 23 wherein said hydrophilic group is a carboxylic acid, a polyether, a polyalcohol, an amine, a sulfonic acid, a phosphonic acid, a phosphonic ester, a phosphate, a phosphate ester, a phosphinic acid, a boronic acid or a borinic acid.
Claim 27
Dependent Claim
224 words
A fluorescent conjugate represented by the formula ##STR33## wherein W, X, Y and Z each independently is --(--CH2) a (CHR 4 )(CH 2 --)-- b ; R, R 1 , R 3 and R 4 each independently is hydrogen, a hydrophilic group or a substituent including a biologically active moiety selected from the group consisting of an antigen, an antibody, a hapten, an Fab fragment and a DNA probe; R 2 is --CO 2 .sup.⊖, SO 3 .sup.⊖ or a substituent including said biologically active moiety; provided that one of R, R 1 , R 2 , R 3 and R 4 is a substituent including said biologically active moiety and at least one of the remaining of R, R 1 , R 3 and R 4 is a hydrophilic group; a and b each independently is an integer of from 0 to 4, provided that the sum of a+b is an integer of from 1 to 4; m and n each is 0 to 1 provided that one of m and n is 0 and the other is 1; p and q each is 0 or 1 provided that one of p and q is 0 and the other is 1; A is a counterion or the counterions to balance the overall charges on the conjugate moiety; and x is 0 or 1.
Claim 28
Dependent Claim
224 words
A fluorescent conjugate represented by the formula ##STR33## wherein W, X, Y and Z each independently is --(--CH2) a (CHR 4 )(CH 2 --)-- b ; R, R 1 , R 3 and R 4 each independently is hydrogen, a hydrophilic group or a substituent including a biologically active moiety selected from the group consisting of an antigen, an antibody, a hapten, an Fab fragment and a DNA probe; R 2 is --CO 2 .sup.⊖, SO 3 .sup.⊖ or a substituent including said biologically active moiety; provided that one of R, R 1 , R 2 , R 3 and R 4 is a substituent including said biologically active moiety and at least one of the remaining of R, R 1 , R 3 and R 4 is a hydrophilic group; a and b each independently is an integer of from 0 to 4, provided that the sum of a+b is an integer of from 1 to 4; m and n each is 0 to 1 provided that one of m and n is 0 and the other is 1; p and q each is 0 or 1 provided that one of p and q is 0 and the other is 1; A is a counterion or the counterions to balance the overall charges on the conjugate moiety; and x is 0 or 1.
Claim 29
Dependent Claim
97 words
The fluorescent conjugate as defined in claim 25 which is represented by the formula ##STR34## wherein R 12 , R 13 , R 14 and R 15 each independently is hydrogen, a hydrophilic group or a substituent including said biologically active moiety, provided that one of R 2 , R 3 , R 12 , R 13 , R 14 and R 15 is a substituent including said biologically active moiety and at least one of the remaining of R 3 , R 12 , R 13 , R 14 and R 15 is a hydrophilic group.
Claim 30
Dependent Claim
19 words
The fluorescent conjugate as defined in claim 26 wherein R 2 is a substituent including said biologically active moiety.
Claim 31
Dependent Claim
97 words
The fluorescent conjugate as defined in claim 25 which is represented by the formula ##STR35## wherein R 12 , R 13 , R 14 and R 15 each independently is hydrogen, a hydrophilic group or a substituent including said biologically active moiety, provided that one of R 2 , R 3 , R 12 , R 13 , R 14 and R 15 is a substituent including said biologically active moiety and at least one of the remaining of R 3 , R 12 , R 13 , R 14 and R 15 is a hydrophilic group.
Claim 32
Dependent Claim
19 words
The fluorescent conjugate as defined in claim 28 wherein R 2 is a substituent including said biologically active moiety.
Claim 33
Dependent Claim
46 words
The fluorescent conjugate as defined in claim 25 wherein said hydrophilic group is a carboxylic acid, a polyether, a polyalcohol, an amine, a sulfonic acid, a phosphonic acid, a phosphonic ester, a phosphate, a phosphate ester, a phosphinic acid, a boronic acid or a borinic acid.
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| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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