Particle-based ligand assay with extended dynamic range
AssigneeBeckman Coulter, Inc.
InventorMichael L. Bell
Life of this patent
Estimating the term…
Abstract
This invention relates to methods of assaying one or more analytes simultaneously. The assays of this invention are capable of providing wide dynamic range and rapid processing times. A wide dynamic working range is achieved by simultaneously incubating a sample which may contain the analyte(s) of interest with two or more independently determinable classes of particles coated with an analyte-specific binding partner. The two or more particle classes differ from each other at least in size. The analyte concentration is obtained from readings derived from these two classes by means of a combined standard curve.
Claims41
A method for assaying an analyte in a sample, comprising:
- a) providing a set of coated particles comprising at least
- i) a first class of particles having a first analyte-specific binding partner immobilized thereon, and
- ii) a second class of particles having a second analyte-specific binding partner immobilized thereon, wherein said first and second classes of coated particles are independently determinable, wherein said particles of said first class of particles are smaller than the particles of said second class, and wherein the first binding partner and the second binding partner are the same;
- b) simultaneously incubating said sample in the presence of said set of particles under conditions that allow the formation of a first complex comprising said analyte bound to said first binding partner immobilized on said first class of particles and/or a second complex comprising said analyte bound to said second binding partner immobilized on said second class of particles;
- c) correlating the amount of said first complex and/or said second complex to the concentration of said analyte in said sample;
- and
- d) determining the concentration of said analyte.
A method for assaying a plurality of analytes in a sample, comprising:
- a) providing a plurality of sets of coated particles equal to the number of analytes to be assayed, wherein each set of coated particles is specific for a particular analyte of interest and is separately determinable, wherein each set comprises at least
- i) a first class of particles having a first binding partner immobilized thereon, and
- ii) a second class of particles having a second binding partner immobilized thereon, wherein said first and second classes of coated particles are independently determinable, wherein said particles of said first class of particles are smaller than the particles of said second class, and wherein the first binding partner and the second binding partner are the same;
- b) simultaneously incubating said sample in the presence of said sets of coated particles under conditions that allow the formation of a set of complexes for each analyte, wherein each set of complexes comprises;
- a first complex comprising said set-specific analyte bound to said first binding partner immobilized on said first class of particles and/or a second complex comprising said set-specific analyte bound to said second binding partner immobilized on said second class of particles;
- and
- c) correlating the amount of said first complex and/or said second complex formed by each set of coated particles to the concentrations of said analytes in said sample;
- and
- d) determining the concentration of said analytes.
A kit for assaying an analyte in a test sample, said kit comprising:
- (a) a set of coated particles comprising at least
- i) a first class of particles having a first analyte-specific binding partner immobilized thereon, and
- ii) a second class of particles having a second analyte-specific binding partner immobilized thereon, wherein said first and second classes of particles are independently determinable, wherein said particles of said first class of particles are smaller than the particles of said second class, and wherein the first binding partner and the second binding partner are the same;
- and
- (b) a labeled ligand, wherein said ligand comprises either a third binding partner for said analyte or an analyte analogue.
Dependencies are inferred from the claim text.