Affinity Separation: M13 Bacteriophage-Activated Superparamagnetic Beads for Affinity Separation

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Title: Affinity Separation: M13 Bacteriophage-Activated Superparamagnetic Beads for Affinity Separation
Authors: Muzard, Julien
Platt, Mark
Lee, Gil U.
Permanent link: http://hdl.handle.net/10197/8386
Date: 6-Aug-2012
Abstract: The growth of the biopharmaceutical industry has created a demand for new technologies for the purification of genetically engineered proteins.The efficiency of large-scale, high-gradient magnetic fishing could be improved if magnetic particles offering higher binding capacity and magnetization were available. This article describes several strategies for synthesizing microbeads that are composed of a M13 bacteriophage layer assembled on a superparamagnetic core. Chemically cross-linking the pVIII proteins to a carboxyl functionalized bead produced highly responsive superparamagnetic particles (SPM) with a side-on oriented, adherent virus monolayer. Also, the genetic manipulation of the pIII proteins with a His6 peptide sequence allowed reversible assembly of the bacteriophage on a nitrilotriacetic acid functionalized core in an end-on configuration. These phage-magnetic particles were successfully used to separate antibodies from high-protein concentration solutions in a single step with a > 90 % purity. The dense magnetic core of these particles makes themfive times more responsive to magnetic fields than commercial materialscomposed of polymer-iron oxide compositesand a monolayer of phage could produced a 1000 fold higher antibody binding capacity. These new bionanomaterials appear to be well-suited to large-scale high-gradient magnetic fishing separation and promise to be cost effective as a result of the self-assembling and self-replicating properties of genetically engineered M13 bacteriophage
Funding Details: Science Foundation Ireland
Type of material: Journal Article
Publisher: Wiley
Copyright (published version): 2012 Wiley
Keywords: Bionanomaterial;M13 filamentous phage;Affinity separation;Downstream processing;Superparamagnetic beads;High gradient magnetic separation
DOI: 10.1002/smll.201290083
Language: en
Status of Item: Peer reviewed
Appears in Collections:Medicine Research Collection

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