AMPLIFICATION OF FLOW-MICROCALORIMETRY SIGNAL BY MEANS OF MULTIPLE BIOAFFINITY LAYERING OF LECTIN AND GLYCOENZYME

This paper demonstrates a positive influence of a special, stepwise te chnique of enzyme immobilization based on the biospecific adsorption o f the glycoenzyme invertase on immobilized concanavalin A (Con A), sub sequent adsorption of the free Con A on the immobilized invertase:Con A support and repeated adsorption of invertase on the support. A 3-fol d repetition of the same procedure designed preliminarily as bioaffini ty layering afforded up to a 10-fold increase in catalytic activity of the immobilized invertase. Reactive hydrogels based on bead cellulose and bead poly(glycidyl methacrylate) were used as immobilization supp orts for the preparation of these highly active preparations. The enha ncement: in catalytic activity of immobilized invertase preparations w as demonstrated thermometrically, by flow microcalorimetry. Further at tractive aspects for utilizing the signal amplification of biosensors with immobilized enzymes are discussed.