Fluorescent conjugates and biological diagnostic assay
AssigneePolaroid Corporation
InventorsMichael J. Arnost, Frank A. Meneghini, Paul S. Palumbo, Stephen G. Stroud
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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.
Claims30
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).sub.a (CHR.sub.4)(CH.sub.2 --)--.sub.b ;
- R, R.sub.1, R.sub.3 and R.sub.4 each independently is hydrogen, a hydrophilic group or a substituent including said analyte or an analogue thereof;
- R.sub.2 is --CO.sub.2.sup..crclbar., SO.sub.3.sup..crclbar. or a substituent including said analyte or an analogue thereof;
- provided that one of R, R.sub.1, R.sub.2, R.sub.3 and R.sub.4 is a substituent including said analyte or an analogue thereof and at least one of the remaining of R, R.sub.1, R.sub.3 and R.sub.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.
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).sub.a (CHR.sub.4)(CH.sub.2 --)--.sub.b ;
- R, R.sub.1, R.sub.3 and R.sub.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.sub.2 is --CO.sub.2.sup..crclbar., SO.sub.3.sup..crclbar. or a substituent including said biologically active moiety;
- provided that one of R, R.sub.1, R.sub.2, R.sub.3 and R.sub.4 is a substituent including said biologically active moiety and at least one of the remaining of R, R.sub.1, R.sub.3 and R.sub.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.
A fluorescent conjugate represented by the formula ##STR33## wherein W, X, Y and Z each independently is --(--CH2).sub.a (CHR.sub.4)(CH.sub.2 --)--.sub.b ;
- R, R.sub.1, R.sub.3 and R.sub.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.sub.2 is --CO.sub.2.sup..crclbar., SO.sub.3.sup..crclbar. or a substituent including said biologically active moiety;
- provided that one of R, R.sub.1, R.sub.2, R.sub.3 and R.sub.4 is a substituent including said biologically active moiety and at least one of the remaining of R, R.sub.1, R.sub.3 and R.sub.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.
Dependencies are inferred from the claim text.