Bacteriophages and Cryogels:A New Efficient Tool in Bioseparation

摘要

An affinity purification procedure for the direct purification of a protein from a complex sample has been developed. The procedure is based on using selected phage clones expressing a peptide with high binding affinity for a selected protein which were covalently coupled to a macroporous monolithic column. Large pore size (10-100 μm) makes it possible to couple phage particles (~ lμm long) as ligands without any risk of blocking the monolithic column. The bound protein was eluted with a purity of >95%. The technique presents a good alternative to conventional immuno-affinity 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. Two case studies are presented: the capture of lactoferrin from defatted milk and of von Willebrand Factor from whole blood or plasma.