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

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Download Small_2012_-_Affinity_Separation_.pdf6.48 MB
Author(s)
Muzard, Julien 
Platt, Mark 
Lee, Gil U. 
Uri
http://hdl.handle.net/10197/8386
Date Issued
06 August 2012
Date Available
02T18:05:48Z March 2017
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
Sponsorship
Science Foundation Ireland
Other Sponsorship
Irish National Institute for Bioprocessing Research and Training (NIBRT)
IEF Marie Curie fellowship
Type of Material
Journal Article
Publisher
Wiley
Journal
Small
Volume
8
Issue
15
Copyright (Published Version)
2012 Wiley
Keywords
  • Bionanomaterial

  • M13 filamentous phage...

  • Affinity separation

  • Downstream processing...

  • Superparamagnetic bea...

  • High gradient magneti...

DOI
10.1002/smll.201290083
Language
English
Status of Item
Peer reviewed
This item is made available under a Creative Commons License
https://creativecommons.org/licenses/by-nc-nd/3.0/ie/
Owning collection
Medicine Research Collection
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