Heterogeneous immunoassay process
First Claim
1. Method for determining an analyte in a liquid sample, comprising:
- (i) contacting said sample to a carrier material comprising (a) a dissolvable labelled immunoreactant which specifically binds to said analyte and which reacts with said analyte to form a complex therebetween and (b) a dissolvable, detectable substance selected from the group consisting of a fluorescent dye and a colored dye which does not participate in the reaction between said analyte and said dissolvable labelled analyte specific immunoreactant and which is soluble in the same solutions and under the same conditions in which said dissolvable labelled specific immunoreactant is soluble wherein (a) and (b) dissolve from said carrier into said liquid sample to form a liquid mixture of said analyte, said labelled immunoreactant and said dissolvable detectable substance,(ii) incubating said liquid mixture with a solid phase which comprises an amount of solid phase bound analyte larger than the amount of said labelled immunoreactant, to form immune complexes between said analyte and said dissolvable labelled immunoreactant and between said solid phase bound analyte and said labelled immunoreactant(iii) separating solid phase bound immune complexes from liquid phase immune complexes;
(iv) measuring the amount of label in the liquid phase to provide a first, uncorrected value indicative of analyte concentration,(v) measuring the amount of said detectable substance in said liquid phase to determine a second measurement value which is proportional to the portion of said labelled immunoreactant dissolved from said carrier which has not formed solid phase immunecomplexes and is present in said liquid phase, and;
(vi) determining a corrected analyte concentration from the difference of said second measurement value from a mean value for said detectable substance and correcting said first uncorrected value by said difference.
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Abstract
The invention teaches a method for determining an analyte in a liquid sample via a heterogeneous assay. The sample is contacted to a carrier material which contains, in addition to one of a labelled analyte analogous substance or a labelled analyte specific substance, a detectable component which does not influence immunological reactions occurring during the course of the assay. A solid phase bound component, which is either an insufficient amount of analyte specific substance or an excess of analyte relative to analyte in the sample is contacted to the sample so as to accomplish formation of solid phase bound complexes. After separation of liquid and solid phase, label is measured in one of these, and the detectable substance is measured in the liquid. Correlation of the values obtained leads to determination of the analyte. Also described is a reagent used in the described method.
16 Citations
6 Claims
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1. Method for determining an analyte in a liquid sample, comprising:
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(i) contacting said sample to a carrier material comprising (a) a dissolvable labelled immunoreactant which specifically binds to said analyte and which reacts with said analyte to form a complex therebetween and (b) a dissolvable, detectable substance selected from the group consisting of a fluorescent dye and a colored dye which does not participate in the reaction between said analyte and said dissolvable labelled analyte specific immunoreactant and which is soluble in the same solutions and under the same conditions in which said dissolvable labelled specific immunoreactant is soluble wherein (a) and (b) dissolve from said carrier into said liquid sample to form a liquid mixture of said analyte, said labelled immunoreactant and said dissolvable detectable substance, (ii) incubating said liquid mixture with a solid phase which comprises an amount of solid phase bound analyte larger than the amount of said labelled immunoreactant, to form immune complexes between said analyte and said dissolvable labelled immunoreactant and between said solid phase bound analyte and said labelled immunoreactant (iii) separating solid phase bound immune complexes from liquid phase immune complexes; (iv) measuring the amount of label in the liquid phase to provide a first, uncorrected value indicative of analyte concentration, (v) measuring the amount of said detectable substance in said liquid phase to determine a second measurement value which is proportional to the portion of said labelled immunoreactant dissolved from said carrier which has not formed solid phase immunecomplexes and is present in said liquid phase, and; (vi) determining a corrected analyte concentration from the difference of said second measurement value from a mean value for said detectable substance and correcting said first uncorrected value by said difference. - View Dependent Claims (2, 3)
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4. Method for determining an analyte in a liquid sample, comprising:
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(i) contacting said sample to a carrier material comprising (a) a dissolvable labelled analyte analogue and (b) a dissolvable, detectable substance selected from the group consisting of a fluorescent dye and a colored dye which is soluble in the same solutions and under the same conditions in which said dissolvable labelled analyte analogue specific immunoreactant is soluble wherein (a) and (b) dissolve in said liquid sample to form a liquid mixture of analyte, labelled analyte analogue and dissolvable detectable substance, (ii) incubating said liquid mixture with a solid phase comprising an amount of a solid phase bound immunoreactant which specifically binds to said analyte, wherein said amount is smaller than the amount of said labelled analyte analogue, to form immune complexes between said analyte and said solid phase bound immunoreactant, wherein said detectable substance dissolved in said liquid does not participate in the reaction between said solid phase bound immunoreactant and one of said analyte and said labelled analyte analogue, (iii) separating said solid phase bound immune complexes from liquid phase immune complexes, (iv) measuring the amount of label in the liquid phase to provide a first, uncorrected value indicative of analyte concentration, (v) measuring the amount of said detectable substance in said liquid phase to determine the portion of said labelled analyte analogue dissolved from said carrier, which has not formed solid phase immune complexes and is present in said liquid phase, and; (v) determining a corrected analyte concentration from the difference of said determination of said second measurement value from a mean value for said detectable substance and correcting said first uncorrected value by said difference. - View Dependent Claims (5, 6)
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Specification