Specific binding assays employing label analog to reduce sample interferences

Patent · Tue Mar 07 04:00:00 EST 1989

OSTI ID:6076751

Ledden, D J; Schroedter, D E

This patent describes a homogeneous apoenzyme reactivation immunoassay method for determining an analyte in a test sample suspected to contain a potentially interfering binding substance. The test sample is combined with a reagent system that includes (i) a labeled reagent comprising a flavin adenine dinucleotide (FAD) label portion and a specific binding portion comprising the analyte or a binding analog thereof, (ii) an antibody capable of binding the analyte, and (iii) apoglucose oxidase, thereby forming a reaction mixture comprising an antibody bound-species and a free-species of the FAD-labeled reagent. The ability of the FAD-label portion of the labeled reagent to combine with the apoglucose oxidase to produce the catalytically active holoenzyme glucose oxidase is measurably different when the FAD-labeled reagent is in the bound-species compared to when in the free-species. The potentially interfering binding substance is capable of binding with the FAD-labeled portion of the labeled reagent. The glucose oxidase activity in the reaction mixture is measured and related to the amount of analyte in the test sample. The improvement consists of adding to the reaction mixture a flavin structural analog of FAD which does not significantly combine with apoglucose oxidase to produce active glucose oxidase but which binds with the potentially interfering FAD-binding substance.

• Assignee: Miles Inc., Elkhart, IN
• Patent Number(s): US 4810635
• OSTI ID: 6076751
• Country of Publication: United States
• Language: English

Related Subjects

550200* -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
ACTIVATION ENERGY
ADENINES
ALDEHYDES
AMINES
ANTIBODIES
ANTIMETABOLITES
AROMATICS
AZAARENES
BINDING ENERGY
BIOCHEMISTRY
CARBOHYDRATES
CHEMICAL ACTIVATION
CHEMICAL BONDS
CHEMISTRY
DRUGS
ENERGY
ENZYME IMMUNOASSAY
ENZYME REACTIVATION
ENZYMES
GLUCOSE
HETEROCYCLIC COMPOUNDS
HEXOSES
IMMUNITY
IMMUNOASSAY
ISOALLOXAZINES
LABELLING
MONOSACCHARIDES
NUCLEOTIDES
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANIC OXYGEN COMPOUNDS
OXIDOREDUCTASES
PURINES
REAGENTS
SACCHARIDES
SAMPLING