Specific binding assay with a prosthetic group as a label compon

Details Specific binding assay with a prosthetic group as a label compon Specific binding assay with a prosthetic group as a label compon

1979-06-04

1980-12-09

Wiseman, Thomas G.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving antigen-antibody binding, specific binding protein...

435177, 435188, C12Q 166, C12N 996, C12N 1102

Patent

active

042385652

ABSTRACT:
A specific binding assay for determining a ligand in a liquid medium employing an organic prosthetic group residue, such as a residue of flavin adenine dinucleotide, flavin mononucleotide, or heme, as a label component in the labeled conjugate. Preferably, the label component is the prosthetic group residue alone or is a holoenzyme residue comprising such prosthetic group residue combined with an apoenzyme in the form of a holoenzyme complex. In the former case, the label component preferably is monitored in the assay by adding an apoenzyme after the binding reaction has been initiated and measuring the resultant holoenzyme activity. In the latter case, the label component is monitored simply by measuring holoenzyme activity. The assay method may follow conventional homogeneous and heterogeneous schemes. Preferred apoenzymes for use in the assay are apoglucose oxidase and apoperoxidase. The assay offers the advantages of colorimetric read-out and of being readily adaptable to automated techniques. Also disclosed are prosthetic group-labeled and enzyme-labeled conjugates for use in the assay, as well as a method for preparing the latter conjugates.

REFERENCES:
patent: 3879262 (1975-04-01), Schuurs
patent: 4040907 (1977-08-01), Ullman et al.
patent: 4134792 (1979-01-01), Boguslaski et al.

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