Journal
SMALL
Volume 8, Issue 15, Pages 2403-2411Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/smll.201200099
Keywords
bionanomaterials; M13 filamentous phage; downstream processing; superparamagnetic beads; high-gradient magnetic separations
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Funding
- Science Foundation Ireland [08/IN/2972]
- Irish National Institute for Bioprocessing Research and Training (NIBRT)
- IEF Marie Curie fellowship [FP7-PEOPLE-2009-IEF 252935]
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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. Chemical cross-linking of the pVIII proteins to a carboxyl-functionalized bead produces 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 allows reversible assembly of the bacteriophage on a nitrilotriacetic-acid-functionalized core in an end-on configuration. These phagemagnetic particles are 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 them five times more responsive to magnetic fields than commercial materials composed of polymer(iron oxide) composites and a monolayer of phage could produce 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.
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