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

• Published in: Process biochemistry (1991) Vol. 48; no. 12; pp. 1972 - 1979
• Main Authors: Urtasun, Nicolás, Baieli, María F., Romasanta, Pablo N., Fernández, Marisa M., Malchiodi, Emilio L., Cascone, Osvaldo, Wolman, Federico J., Miranda, María V.
• Format: Journal Article
• Language: English
• Published: Elsevier Ltd 01.12.2013
• Subjects: Adsorption; antibacterial properties; Antiinfectives and antibacterials; antimicrobial peptides; antiparasitic agents; Baculovirus; Bovine lactoferricin; cattle; chromatography; culture media; Dye affinity chromatography; Dyes; pathogens; Peptides; Proteins; Purification; Recombinant; salt concentration; Surface chemistry; Surface plasmon resonance; triazines; Dye affinity chromatography; Surface plasmon resonance; Baculovirus; Bovine lactoferricin
• ISSN: 1359-5113; 1873-3298
• DOI: 10.1016/j.procbio.2013.07.018

•SPR was used to test five triazine dyes for Lfcin B affinity interaction.•Red HE-3B and Yellow HE-4R dyes were selected and immobilized on Sepharose.•Lfcin B was expressed as a fusion protein with GST by using BEVS.•Yellow HE-4R dye appeared as a new ligand to purify Lfcin B.•We showed a novel application for SPR as a tool to select dye affinity ligands. 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. 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.