Immobilized enzymes and cells in poly(N-vinyl caprolactam)-based hydrogels- Preparation, properties, and applications in biotechnology and medicine

A one-step mild method for entrapping animal cells and enzymes in macroporo us composite poly(N-vinyl caprolactam)-calcium alginate (PVCL-CaAlg) hydrog els is described. Some properties of immobilized enzymes, such as thermal a nd storage stabilities and stability in water/organic media were investigat ed. Composite PVCL-CaAlg gels were successfully applied to immobilize a num ber of proteases, namely, trypsin, alpha-chymotrypsin, carboxypeptidase B, and thrombin. Thermal stability of the immobilized preparations obtained by entrapment in hydrogel beads allowed us to use them at 65-80 degrees C, wh ile the native enzymes were completely inactivated at 50-55 degrees C. Vari ous application's of enzymes and cells immobilized in beads were demonstrat ed. Immobilized trypsin and carboxypeptidase B were applied to prepare huma n insulin from recombinant proinsulin. The hydrogel beads with entrapped al pha-chymotrypsin were used in enantioselective hydrolysis of Shiff's base o f D,L-phenylalanine ethyl ester (SBPH) in acetonitrile/water medium. Thromb in immobilized in PVCL-based hydrogel films was shown to be a promising com pound for wound treatment. To prepare pure preparations of monoclonal antib odies (MAb) several hybridoma cell lines, including hybridoma cell lines pr oducing MAb to interleukin-2, were successfully cultivated in the hydrogel beads.