Fluorescent labels and labeled species and their use in analytical elements and determinations
First Claim
1. A fluorescent labeled physiologically reactive species comprisinga conjugate of a physiologically reactive species bound to a polysaccharide,said conjugate bound through said polysaccharide to a polymeric particle which is derived from a loadable latex having a discontinuous phase which consists essentially of a polymer prepared from one or more ethylenically unsaturated polymerizable monomers, and an aqueous continuous phase, said polymeric particle having incorporated therein a fluorescent rare earth chelate.
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Abstract
Fluorescent labels comprise a polysaccharide bound to a polymeric particle which contains a fluorescent rare earth chelate. These labels can be attached to any of a variety of physiologically reactive species to provide labeled species which have improved stability in aqueous solutions. The labeled species are particularly useful in specific binding assays to determine an immunologically reactive ligand, e.g. a hapten, in either solution or dry analytical techniques.
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Citations
18 Claims
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1. A fluorescent labeled physiologically reactive species comprising
a conjugate of a physiologically reactive species bound to a polysaccharide, said conjugate bound through said polysaccharide to a polymeric particle which is derived from a loadable latex having a discontinuous phase which consists essentially of a polymer prepared from one or more ethylenically unsaturated polymerizable monomers, and an aqueous continuous phase, said polymeric particle having incorporated therein a fluorescent rare earth chelate.
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6. A fluorescent labeled specific binding ligand analog for determining a specific binding ligand in an aqueous liquid, said labeled analog comprising
a conjugate of said ligand bound to a polysaccharide, said conjugate bound through said polysaccharide to a polymeric particle which is derived from a loadable latex having a polymeric discontinuous phase which consists essentially of a polymer comprising (a) from 0 to 100 weight percent of units derived from a substituted or unsubstituted styrene or vinyl naphthyl, (b) from 0 to about 90 weight percent of units derived from an ethylenically unsaturated monomer of the structure ##STR8## wherein R is hydrogen or alkyl, R1 is hydrogen or methyl and R2 is hydrogen, halogen, alkyl, cyano, ##STR9## wherein R3 is hydrogen or R4, and R4 is an aliphatic group, and (c) from 0 to about 10 weight percent of units derived from an ethylenically unsaturated monomer containing one or more sulfonic acid or carboxy groups, or an ammonium or alkali metal salt thereof, and an aqueous continuous phase, said polymeric particle having incorporated therein a fluorescent rare earth chelate.
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8. A dry analytical element comprising an absorbent carrier material, and a fluorescent labeled physiologically reactive species,
said labeled species comprising a conjugate of a physiologically reactive species bound to a polysaccharide, said conjugate bound through said polysaccharide to a polymeric particle which is derived from a loadable latex having a discontinuous phase which consists essentially of a polymer prepared from one or more ethylenically unsaturated polymerizable monomers, and an aqueous continuous phase, said polymeric particle having incorporated therein a fluorescent rare earth chelate.
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10. A dry analytical element for the determination of a specific binding ligand in an immunoassay, said element comprising
a support having thereon a porous spreading zone, and a fluorescent labeled ligand analog of said ligand, said labeled analog comprising a conjugate of said ligand bound to a polysaccharide, said conjugate bound through said polysaccharide to a polymeric particle which is derived from a loadable latex having a discontinuous phase which consists essentially of a polymer, and an aqueous continuous phase, said polymer comprising (a) from 40 to 100 weight percent of units derived from a substituted or unsubstituted styrene or vinyl naphthyl, and (b) 0 to about 60 weight percent of units derived from an ethylenically unsaturated monomer of the structure ##STR10## wherein R is hydrogen or alkyl, R1 is hydrogen or methyl, and R2 is hydrogen, halogen, alkyl, cyano, ##STR11## wherein R3 is hydrogen or R4, and R4 is an aliphatic group, said polymeric particle having incorporated therein a fluorescent rare earth chelate.
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14. A method for the determination of a specific binding ligand in an aqueous liquid, said method comprising the steps of:
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A. in the presence of a receptor for said ligand, contacting a sample of said liquid with a fluorescent labeled specific binding ligand analog, said ligand analog comprising a conjugate of said ligand bound to a polysaccharide, said conjugate bound through said polysaccharide to a polymeric particle which is derived from a loadable latex having a discontinuous phase which consists essentially of a polymer prepared from one or more ethylenically unsaturated polymerizable monomers, and an aqueous continuous phase, said polymeric particle having incorporated therein a fluorescent rare earth chelate to form a complex between said receptor and said ligand analog, B. separating said complex from uncomplexed ligand analog, and C. fluorometrically detecting either said complexed or uncomplexed ligand analog as a measure of said ligand. - View Dependent Claims (15, 16, 17)
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18. A dignostic test kit for the determination of a specific binding ligand, said kit comprising
(i) a receptor for said ligand and (ii) a fluorescent labeled specific binding ligand analog comprising a conjugate of said ligand bound to a polysaccharide, said conjugate bound through said polysaccharide to a polymeric particle which is derived from a loadable latex having a discontinuous phase which consists essentially of a polymer prepared from one or more ethylenically unsaturated polymerizable monomers, and an aqueous continuous phase, said polymeric particle having incorporated therein a fluorescent rare earth chelate.
Specification