Use of Periodate Method for the Preparation of Steroid Enzyme Conjugate through Adipic Acid Dihydrazide as Linker and its Use in Immunoassy

Horseradish peroxidase (HRP), a glycoenzyme, is the most widely used enzyme in enzyme immunoassy. Poor availability of amino groups in HRP has been observed, which is attributed by the change in extraction condition for its isolation from the ischolar_mains of horseradish plant. There by it is difficult to couple immunoreactants to HRP directly by bi-functional coupling reagents. To overcome this obstacle, the present conjugation procedure exploit periodate oxidation of carbohydrate moiety of HRP and subsequently formation of hydrazone bond between newly formed HRP aldehyde and one hydrazide group of adipic acid dihydrazide (ADH).