Radiation and photografting as complementary techniques for immobilizing bioactive materials

The technique of radiation grafting to immobilize a typical enzyme, trypsin, to three representative backbone polymers namely polypropylene, PVC and polystyrene, is described. The process involves the simultaneous grafting of p-nitrostyrene to the polymer, the nitro group subsequently being converted to its isothiocyanato derivative to which trypsin is attached. For this immobilization work, the utilization of additives which enhance grafting is shown to be an advantage. Typical of the additives used are mineral acids (approx. 0.1M) and polyfunctional monomers (approx. 1% v/v), the two acting in a synergistic fashion when used together. A new class of additives, namely metal salts such as LiC104, which also increase grafting yields has now been discovered. The reactivity of these salts in grafting enhancement is compared with that of mineral acid. A new model for the modus operandi of metal salts as well as acids in grafting enhancement is proposed whereby increased grafting yields are attributed to increased partitioning of monomer into the graft region in the presence of ionic solutes. The significance of using these additives to prepare radiation copolymers for enzyme immobilization is discussed. In addition to ionizing radiation, UV is also shown to be of value as an initiator for the preparation of graft copolymers for enzyme immobilization, however the presence of sensitizer in the copolymer may limit the photografting method. The possibility of using radiation curing processes involving electron beam (EB) and high powered (300 W/inch) UV sources for immobilization work has been explored and found to be feasible. The curing system possesses great potential for the current immobilization work since complete polymerization is achieved in a fraction of a second. (author)