IONTOSORB - Bead Cellulose Derivatives
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Immobilization Enzymes by Covalent Bond

Immobilization of Enzymes by Ugi's Reaction

Ugi's reaction [Ugi I.:  Angew. Chem.,  74 (1962) 9] is often used as a immobilization method and therefore it was also used for sorbents IONTOSORB DETA and IONTOSORB AV. The procedure is described as follows :

To 1 g support 10 ml of the enzyme solution was added. The enzyme solution contained GOD, KAT - 10 mg in 10 ml phosphate buffer (c = 0.1 mol/l) at pH 7.2 or CHT - 20 mg in 10 ml phosphate buffer (c = 0.1 mol/l) at pH 7.8. After that 60 µl acetic acid (c = 0.5 mol/l), 3 µl glutaraldehyde (25 %) and 30 µl cyclohexylisocyanide were added. The mixture was shook 20 hours at 4 ^oC. The obtained conjugate was rinsed in the alternate way with phosphate buffer and NaCl solution in the same buffer with the adequate pH value. Immobilized enzymes were covered with phosphate buffer and saved at 4 ^oC.

alpha-Chymotrypsine (CHT), Glucoseoxidase (GOD) and Catalase (KAT)

Results of binding are given in the following table:

MS = moist support                                 DS = dry support