Specific binding assay employing an enzyme-cleavable...

C - Chemistry – Metallurgy – 12 – Q

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167/43

C12Q 1/34 (2006.01) G01N 33/54 (1980.01)

Patent

CA 1148080

ABSTRACT An improved homogeneous specific binding assay method for determining a ligand in a liquid medium, wherein a reaction mixture is formed by combining the liquid medium with reagent means, including a conjugate having a label component and a binding component, to form a binding reaction system having a bound-species and a free-species of the labeled conjugate, the label component of the conjugate comprising an enzyme substrate- active portion and an indicator portion whereby the conjugate is cleavable by an enzyme to produce a detectable indicator product, the labeled conjugate being substantially inactive as a substrate for the enzyme when in the bound-species, and wherein the cleaving enzyme is added to the reaction mixture and the resulting indicator product measured as a function of the amount of the ligand in said liquid medium. The improvement comprises employing as the label component of the conjugate, a residue having the formula: Image wherein R is a linking group through which the residue is covalently bound to the binding component, and employing .beta.-galactosidase as the cleaving enzyme whereby the .beta.-galactosyl group in the residue can be cleaved to release a detectable dye product.

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