Assays for macromolecular analytes
AssigneeSiemens Healthcare Diagnostics Inc.
InventorJames E. Duffy
Life of this patent
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Abstract
A method of determining a macromolecular analyte in a sample suspected of containing the macromolecular analyte is disclosed. The sample and a conjugate reagent comprising a small molecule and a binding partner for the macromolecular analyte are combined in a medium. The conjugate reagent and a labeled binding partner for the small molecule are combined. The conjugate reagent or the medium is examined for an amount of labeled binding partner for the small molecule that is bound to the small molecule, which is related to the amount of the macromolecular analyte in the sample.
Claims22
A method of determining a macromolecular analyte in a sample suspected of containing the macromolecular analyte, the method comprising:
- (a) combining in a medium the sample and a conjugate reagent comprising a small molecule and a binding partner for the macromolecular analyte,
- (b) combining the conjugate reagent and a labeled binding partner for the small molecule, and
- (c) examining the conjugate reagent or the medium for an amount of labeled binding partner for the small molecule that is bound to the small molecule and relating the amount thereof to the amount of the macromolecular analyte in the sample,
- wherein said macromolecular analyte, when present, binds to the binding partner for the macromolecular analyte and has a size or a shape or a combination of both size and shape that hinders the ability of the binding partner for the small molecule to bind to the small molecule when the small molecule is part of the reagent that also comprises the binding partner that is specific for the macromolecular substance, wherein the macromolecular analyte has a molecular weight of at least 5000 and is selected from the group consisting of antibodies, enzymes, hormones, polynucleotides, and carbohydrates, and
- wherein the small molecule is an organic molecule that has a molecular weight in the range of from 100 to 2000, comprises functional groups that are reactive with the binding partner for the macromolecular analyte and is bound to the binding partner for the macromolecular analyte.
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The method according to claim 3 or 21, wherein the support is a non-magnetic particle, a magnetic particle, a plate, or a tube.
The method according to claim 1 or 21, wherein the small molecule is selected from the group consisting of biotin, desthiobiotin, dinitrophenol, digoxin, digoxigenin, rhodamine, and fluorescein.
The method according to claim 1 or 21, wherein the label of the labeled binding partner is selected from the group consisting of enzymes, fluorescers, chemiluminescers, bioluminescers, electroluminescers, sensitizers, and radioactive materials.
The method according to claim 3 or 21, wherein the support comprises a label wherein the label is interactive with the label of the labeled binding partner.
The method according to claim 1 or 21, wherein the sample is a body excretion, body aspirant, body excisant or body extractant.
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The method according to claim 1 or 21, wherein the method is a homogeneous assay method.
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The method according to claim 22, wherein the labeled binding partner is labeled with a particle comprising a sensitizer label and the particle having bound thereto a small molecule comprises a chemiluminescent label.
The method according to claim 22, wherein the particle having bound thereto a small molecule is a magnetic particle.
The method according to claim 22, wherein the small molecule is bound to the antigen, which is bound to the particle.
Dependencies are inferred from the claim text.