Activated apoglucose oxidase, method of preparing it, its use in specific binding assay methods and reagent means and test kits containing it

摘要

A method for increasing the ability of apoglucose oxidase to combine with flavin adenine dinucleotide (FAD) and derivatives thereof to form active glucose oxidase by interacting apoglucose oxidase with an immunologically derived binding substance, e.g., an antibody or a fragment thereof, having a specific binding affinity for glucose oxidase. The apoglucose oxidase/anti-glucose oxidase immune complex is characterized by enhanced ability to combine with FAD and FAD-derivatives to yield glucose oxidase activity. The activation is particularly significant at temperatures elevated from room temperature, e.g., between 30°-45°C. An improved homogeneous specific binding assay method is provided for determining ligands wherein an FAD label is used and is monitored by its ability to combine with apoglucose oxidase to form active glucose oxidase by including anti-glucose oxidase in the reaction mixture.