PROCESS FOR PREPARING THE ENZYMATIC MARKER TESTOSTERONE-3-CARBOXYMETHYLOXIM ALKALINE PHOSPHATASE

摘要

The invention relates to a process for preparing the enzymatic marker testosterone-3-carboxymethyloxim alkaline phosphatase, which may be used in ELISA technique of dosing the testosterone hormone in biological liquids. According to the invention, the process consists in preparing testosterone-3-carboxymethyloxim alkaline phosphatase from testosterone and oxy aminoacetic acid, dissolving then 250 parts by weight of testosterone-3-carboxymethyloxim in 1 part by volume of dioxane, mixing then with 10 parts by volume of tributhylamine and 5 parts by volume chloroformiate solution in dioxane medium, stirring the mixture for 30 min., followed by coupling approximately 2,500 U /mg covalently with alkaline phosphatase enzyme of specific activity, purifying next with Sephadex LH-20, having as elution solvent a mixture of ethylic alcohol : water 1: 5 (V/V), subjecting to lyophilization and depositing at a temperature 4 degree C, in order to be used.