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

NICOLÁS URTASUN; MARÍA FERNANDA BAIELI; PABLO N. ROMASANTA; MARISA M. FERNÁNDEZ; EMILIO L. MALCHIODI; OSVALDO CASCONE; FEDERICO JAVIER WOLMAN; MARÍA VICTORIA MIRANDA

PROCESS BIOCHEMISTRY - (Print)

ELSEVIER SCI LTD

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

1359-5113

Bovine lactoferricin (Lfcin B) belongs to the antimicrobial peptide family, which is thefirst 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. Recombinant Lfcin B was expressed as a fusion protein with GST (Lfcin B-GST) by using the baculovirus expression vector system and the dye-Sepharose matrices were assayed for Lfcin B-GST adsorption and subsequent elution. Red HE-3B and Yellow HE-4R dyes were selected and immobilized on a Sepharose-4B matrix for further purification studies. 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