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

NICOLAS URTASUN; FERNANDA BAIELI; PABLO ROMASANTA; MARISA M. FERNANDEZ; EMILIO MALCHIODI; OSVALDO CASCONE; FEDERICO J. WOLMAN; MARÍA V. MIRANDA

PROCESS BIOCHEMISTRY

ELSEVIER

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

0032-9592

Bovine lactoferricin (Lfcin B) belongs to the antimicrobial peptide family, which is the first line of defense against pathogens in many organisms. Lfcin B has important applications due to its antiviral, antifungal, antiparasitic, anticancer/tumor and antibacterial activity. In this work, we tested five triazine dyes for Lfcin B affinity interactions using Surface Plasmon Resonance (SPR) technology. Red HE-3B and Yellow HE-4R dyes were selected and immobilized on a Sepharose-4B matrix for further purification studies. Recombinant Lfcin B was expressed as a fusion protein with Glutathione -S- Transferase (Lfcin B-GST) by using baculovirus expression vector system and the dye-Sepharose matrices were assayed for Lfcin B-GST adsorption and subsequent elution. The Yellow HE-4R-Sepharose matrix was specific for Lfcin B and allowed adsorption of Lfcin B-GST directly from the culture medium even at high salt concentration. This novel application of SPR to screen possible dye-peptide interactions could be relevant to purify other peptides or proteins by using low-cost dye-affinity chromatography.