Activation of poly[N-(2-hydroxypropyl)methacrylamide] for the binding of bioactive molecules, 2. Activation with cyanogen bromide

The amount of chymotrypsinogen linked to poly[N-(2-hydroxypropyl)methacrylamide], which was activated by cyanogen bromide, under various conditions was determined. The optimal activation time was found to be indicated by the beginning of an invariable base consumption during the reaction of the polymer with cyanogen bromide. The degree of activation increased with the concentration of cyanogen bromide and with the pH, and the yield of coupling can be increased by variation of the temperature, concentration of the reaction components, and pH. At room temperature and pH 9,3, the coupling reaction was completed in less than 24 h. The stability of the activated polymer depends strongly on temperature and pH. The analysis of the active moieties on the polymer carrier revealed that the application of the generally accepted mechanism for cyanogen bromide activation of polysaccharide carriers to the activation of poly[N-(2-hydroxypropyl)methacrylamide] remains to be proved.