Desarrollo de una matriz de pseudobioafinidad de bajo costo para la purificación de lactoferrina

WOLMAN, FEDERICO JAVIER; NAVARRO DEL CAÑIZO, AGUSTÍN ANDRÉS; GONZÁLEZ MAGLIO, DANIEL; CASCONE, OSVALDO

Barcelona, España

Congreso; 14th European Congress on Biotechnology; 2009

European Federation of Biotechnology

The main challenge of downstream processing of proteins is to have chromatographic matrices that optimise the performance/cost ratio. Bovine lactoferrin (Lf) was taken as the model as it is a natural protein with antimicrobial properties. Macrobeads of cross-linked chitosan were sinthesised and derivatised with the dye Red HE-3B. Two cross-link reagents at different concentrations were assayed: epichlorhidrin (E) and glutaraldehyde (G). Maximum capacity and dissociation constant for Lf adsorption to the epichlorhidrin cross-linked beads were 21.4 and 0.81 mg/g respectively. For glutaraldehyde cross-linked beads, the values were 21.4 and 0.51 mg/g respectively. Lf elution was maximal with 2 M sodium chloride in 25 % ethyleneglycol at pH 7 for both systems, being higher for epichlorhidrin cross-linked beads (60 % vs 25 %). For Lf purification from bovine colostrum, epichlorhidrin cross-linked beads showed a better performance: at the same sample:matrix ratio, they adsorbed all the lactoferrin contained in the sample while the glutaraldehyde cross-linked beads only adsorbed 80 % of the Lf contained in the sample.