Triazinic dye ligand selection by surface plasmon resonance for recombinant lactoferricin purification.

URTASUN N; BAIELI MF; ROMASANTA P; FERNÁNDEZ M; MALCHIODI E; CASCONE O; WOLMAN F; MIRANDA MV

PROCESS BIOCHEMISTRY

ELSEVIER

Lugar: Amsterdam; Año: 2013 vol. 48 p. 1972 - 1979

0032-9592

2AbstractBovine lactoferricin (Lfcin B) belongs to the antimicrobial peptide family, whichis the first line of defense against pathogens in many organisms. Lfcin B has importantapplications due to its antiviral, antifungal, antiparasitic, anticancer/tumor andantibacterial activity. In this work, we tested five triazine dyes for Lfcin B affinityinteractions using Surface Plasmon Resonance (SPR) technology. Red HE-3B andYellow HE-4R dyes were selected and immobilized on a Sepharose-4B matrix forfurther purification studies. Recombinant Lfcin B was expressed as a fusion proteinwith GST (Lfcin B-GST) by using baculovirus expression vector system (BEVS) andthe dye-Sepharose matrices were assayed for Lfcin B-GST adsorption and subsequentelution. The Yellow HE-4R-Sepharose matrix was specific for Lfcin B and allowedadsorption of Lfcin B-GST directly from the culture medium even at high saltconcentration. This novel application of SPR to screen possible dye-peptide interactionscould be relevant to purify other peptides or proteins by using low-cost dye-affinitychromatography.