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US 7,029,856 B1Granted
Methods for determination of an analyte
AssigneeDade Behring Marburg GmbH
InventorThomas Wissel
Life of this patent
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Abstract
The invention relates to analytical methods in which the partition of a labeled substance between a liquid and a solid phase is determined. The assays include solid-phase reagents which can be particulate or monolithic such as, for example, a coated tube. Assays of this type are known per se to the person skilled in the art and include immunoassays and immunometric assays.
Claims9
A heterogeneous immunochemical method for the determination of an analyte in a sample in a liquid medium, comprising:
- a) incubating the sample in a reaction vessel with a particulate solid-phase on which a first binding partner which specifically binds to the analyte is immobilized and with a second binding partner which specifically binds to the analyte, wherein a fluorescent or chemiluminescent label is conjugated to the second binding partner, and wherein after addition of the solid-phase and the second binding partner the solid-phase is suspended in the liquid medium, whereby the second binding partner is partitioned between the liquid medium and the particulate solid-phase and the solid-phase is subjected to magnetic or gravitational attraction to attract the solid-phase in the liquid medium to an interior surface of the reaction vessel;
- b) removing the first liquid medium from the solid-phase prior to adding a second liquid medium to the solid-phase;
- c) resuspending the solid-phase in the second liquid medium;
- d) adding a reagent A, which causes a separation of the solid-phase and the label by elimination of at least one intermolecular linkage which is essentially composed of hydrogen bonds and ionic, hydrophobic and van der Waals interactions;
- e) removing the solid-phase by magnetic or gravitational attraction;
- and
- f) determining the amount of analyte in the sample, wherein the reagent A contains in aqueous solution one substance selected from the group of reagents consisting of:
- a chaotropic agent, and
- a buffer for adjusting the pH to a range of 1.5 to 3;
- wherein a) through f) are performed in sequential order.
7 more elements
A heterogeneous immunochemical method for the determination of an analyte in a sample in a liquid medium, comprising:
- a) incubating the sample in a reaction vessel with a particulate solid-phase on which a first binding partner which specifically binds to the analyte is immobilized and with a second binding partner which specifically binds to the analyte, wherein a fluorescent or chemiluminescent label is conjugated to the second binding partner, and wherein after addition of the solid-phase and the second binding partner the solid-phase is suspended in the liquid medium, whereby the second binding partner is partitioned between the liquid medium and the particulate solid-phase and the solid-phase is subjected to magnetic or gravitational attraction to attract the solid-phase in the liquid medium to an interior surface of the reaction sample;
- removing the first liquid medium from the solid-phase prior to adding a second liquid medium to the solid-phase;
- c) resuspending the solid-phase in the second liquid medium;
- d) adding a reagent A, which causes a separation of the solid-phase and the label by elimination of at least one intermolecular linkage which is essentially composed of hydrogen bonds and ionic, hydrophobic and van der Waals interactions;
- e) removing the solid-phase by magnetic or gravitational attraction;
- and
- f) determining the amount of analyte in the sample, wherein the reagent A contains in an aqueous solution one substance selected from the group of reagents consisting of:
- KSCN,
- guanidinium hydrochloride,
- NaClO4,
- urea,
- propionic acid,
- dimethylformamide, and
- glycine for adjusting the pH to a range of 1.5 to 3;
- wherein a) through 0 are performed in sequential order.
12 more elements
Dependencies are inferred from the claim text.