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

Paper ID Volume ID Publish Year Pages File Format Full-Text
34429 45025 1979 8 PDF Available
Title
Triazinic dye ligand selection by surface plasmon resonance for recombinant lactoferricin purification
Abstract

•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.

Keywords
Baculovirus; Bovine lactoferricin; Dye affinity chromatography; Surface plasmon resonance
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Triazinic dye ligand selection by surface plasmon resonance for recombinant lactoferricin purification
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Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 48, Issue 12, December 2013, Pages 1972–1979
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us