title: Immobilised peptide displaying phages as affinity ligands. Purification of lactoferrin from defatted milk.
abstract: An affinity purification procedure for the direct purification of lactoferrin from defatted (skimmed) milk has been developed. The procedure is based on using selected phage clones expressing a peptide with high binding affinity for lactoferrin which were covalently coupled to macroporous poly(dimethylacrylamide) monolithic column. Large pore size (10-100 microm) of macroporous poly(dimethylacrylamide) makes it possible to couple long (1 microm) phage particles as ligands without any risk of blocking the monolithic column. Bound lactoferrin was eluted using 1M NaCl with a purity of >95%. The technique presents a good alternative to conventional immunoaffinity chromatography for purification of a protein of interest from complex samples due to (i) the robustness of the system in terms of recovery and ligand leakage and (ii) economical aspect in terms of low ligand cost.
authors.affiliation: Interdisciplinary Research Center, Katholieke Universiteit Leuven Campus Kortrijk, E. Sabbelaan 53, 8500 Kortrijk, Belgium. wim.noppe@kulak.ac.be
authors.forename: Wim
authors.initials: W
authors.lastname: Noppe
authors.forename: Fatima M
authors.initials: FM
authors.lastname: Plieva
authors.forename: Igor Yu
authors.initials: IY
authors.lastname: Galaev
authors.forename: Karen
authors.initials: K
authors.lastname: Vanhoorelbeke
authors.forename: Bo
authors.initials: B
authors.lastname: Mattiasson
authors.forename: Hans
authors.initials: H
authors.lastname: Deckmyn
mesh_terms: D000178:Acrylamides / Q000737:chemistry; D002846:Chromatography, Affinity / Q000295:instrumentation / Q000379:methods*; D005260:Female; D006801:Humans; D007781:Lactoferrin / Q000302:isolation & purification*; D008024:Ligands; D008895:Milk, Human / Q000737:chemistry*; D019151:Peptide Library
chemical_list: D000178:Acrylamides; D008024:Ligands; D019151:Peptide Library; C429790:poly(N,N-dimethylacrylamide); D007781:Lactoferrin