Optimization of glutaraldehyde activation of a support for enzyme immobilization

The influence of several parameters (support porosity, glutaraldehyde concentration, time of action, pH) on the activation reaction of an amine porous silica (Spherosil) with glutaraldehyde has been studied. Glutaraldehyde binding onto the support was followed by measuring the carbon content of the activated silica.We established comparisons between the quantity of glutaraldehyde retained on the support after activation and the capacity of the activated silica to bind trypsin. We have defined the optimal conditions for Spherosil activation and prepared derivatives with high enzymatic activity.Our results are in agreement with a reaction mechanism with glutaraldehyde in a polymeric form resulting from aldol condensation, rather than in a monomeric form.