Properties of human parotid amylase immobilized by covalent binding to glass or sepharose or by reaction with immobilized antibody

Human parotid amylase was immobilized by covalent binding to CNBr-activated Sepharose, to Corning GAO-3940 silica glass biomaterial support by the diazonium reaction or reaction with glutaraldehyde, or as a result of the antigen-antibody reaction between rabbit antihuman parotid amylase IgG that was covalently bonded to GAO glass and soluble amylase. The amylase directly bonded to the supports showed constant activity at flow rates of 3-15 ml/min through a 1.76-cm³ (8-mm diameter) support bed, did not lose enzyme into a circulating starch solution, retained its activity in the presence of soluble antiamylase IgG, was optimally active at 35°-40°C, and lost activity at 40°-45°C. When the enzyme was bound by interaction with immobilized antibody, full activity was expressed, but some enzyme was solubilized by a circulating starch solution. Immobilization of either amylase or antiamylase IgG makes dissolution of the antigen-antibody bond difficult.