Pseudobiospecific chromatographic convective matrices versus traditional soft-gel diffusion systems for protein purification

WOLMAN FEDERICO J.; IANNUCCI NANCY B.; MARANI MARIELA M.; GRASSELLI MARIANO; CASCONE OSVALDO

San Carlos de Bariloche, Neuquen, ARGENTINA

Congreso; SAIB XXXIX Reunión Anual (Sociedad Argentina de Investigación Bioquímica y Biología Molecular); 2003

SAIB

Taking into account that up to 80 % of the biotechnological production costs of a protein is due to recovery and purification processes, development of integrative methods of high capacity, resolution and productivity is desirable. The utilisation of pseudobiospecific ligands such as triazinic dyes immobilised on macroporous membranes such as hollow fibres becomes an interesting alternative strategy for lowering the purification costs. Membranes were activated by direct grafting of monomers with gamma radiation and the triazinic dye Cibacron Blue F3G-A was immobilised prior epoxy group opening with ammonia. Different monomer mixtures were grafted on the membranes (GMA:DMAA in 1:1, 1:2, 1:3). A cartridge was built with 8 fibres of 12.7 cm lenght using those having the best hydrodynamic properties (GMA:DMAA 1:3). The comercial proteolytic preparation Flavourzyme- composed of 4 main proteins, but only one displaying proteolytic activity- was chosen as study sample. The neutral protease from Flavourzyme showed affinity for immobilised Cibacron. As compared with the standard Cibacron-Sepharose matrix, productivity was raised from 318 UE/h to 2180 UE/h working at the maximum allowed volumetric flux (1 ml/min) and at 5 ml/min respectively.