M13 bacteriophage-activated superparamagnetic beads for affinity separation.

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 His(6) peptide sequence allows reversible assembly of the bacteriophage on a nitrilotriacetic-acid-functionalized core in an end-on configuration. These phage-magnetic 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.

[1]  A. Belcher,et al.  Genetically Driven Assembly of Nanorings Based on the M13 Virus , 2004 .

[2]  John McCafferty,et al.  Beyond natural antibodies: the power of in vitro display technologies , 2011, Nature Biotechnology.

[3]  John M. Walker,et al.  The Proteomics Protocols Handbook , 2005, Humana Press.

[4]  K. Avery,et al.  M13 Bacteriophage as a Biological Scaffold for Magnetically-Recoverable Metal Nanowire Catalysts: Combining Specific and Nonspecific Interactions To Design Multifunctional Nanocomposites , 2009 .

[5]  Abhinav A Shukla,et al.  Recent advances in large-scale production of monoclonal antibodies and related proteins. , 2010, Trends in biotechnology.

[6]  Phage matrix for isolation of glioma cell membrane proteins. , 2004, BioTechniques.

[7]  V. Petrenko,et al.  Phages from landscape libraries as substitute antibodies. , 2000, Protein engineering.

[8]  Ki Tae Nam,et al.  Stamped microbattery electrodes based on self-assembled M13 viruses , 2008, Proceedings of the National Academy of Sciences.

[9]  G. P. Smith,et al.  A library of organic landscapes on filamentous phage. , 1996, Protein engineering.

[10]  Anna Merzlyak,et al.  Phage as templates for hybrid materials and mediators for nanomaterial synthesis. , 2006, Current opinion in chemical biology.

[11]  Chuanbin Mao,et al.  Self-assembly of drug-loaded liposomes on genetically engineered target-recognizing M13 phage: a novel nanocarrier for targeted drug delivery. , 2009, Small.

[12]  W. Scott,et al.  Molecular structure of fd (f1, M13) filamentous bacteriophage refined with respect to X-ray fibre diffraction and solid-state NMR data supports specific models of phage assembly at the bacterial membrane. , 2006, Journal of molecular biology.

[13]  Xiaodi Su,et al.  QCM-D analysis of binding mechanism of phage particles displaying a constrained heptapeptide with specific affinity to SiO2 and TiO2. , 2006, Analytical chemistry.

[14]  K. Schulten,et al.  Molecular biomimetics: nanotechnology through biology , 2003, Nature materials.

[15]  J. M. Harris,et al.  Introduction to Biotechnical and Biomedical Applications of Poly(Ethylene Glycol) , 1992 .

[16]  Brian D. Kelley,et al.  DOWNSTREAM PROCESSING OF MONOCLONAL ANTIBODIES: CURRENT PRACTICES AND FUTURE OPPORTUNITIES , 2008 .

[17]  Dong Soo Yun,et al.  Cobalt ion mediated self-assembly of genetically engineered bacteriophage for biomimetic Co-Pt hybrid material. , 2006, Biomacromolecules.

[18]  U. Bläsi,et al.  Genetically modified filamentous phage as bactericidal agents: a pilot study , 2003, Letters in applied microbiology.

[19]  S. Deutscher,et al.  Melanoma imaging with pretargeted bivalent bacteriophage. , 2007, Journal of nuclear medicine : official publication, Society of Nuclear Medicine.

[20]  George Georgiou,et al.  Virus-Based Toolkit for the Directed Synthesis of Magnetic and Semiconducting Nanowires , 2004, Science.

[21]  V. Petrenko,et al.  Landscape phage fusion protein-mediated targeting of nanomedicines enhances their prostate tumor cell association and cytotoxic efficiency. , 2010, Nanomedicine : nanotechnology, biology, and medicine.

[22]  Ahmad S. Khalil,et al.  Single M13 bacteriophage tethering and stretching , 2007, Proceedings of the National Academy of Sciences.

[23]  E. Goldman,et al.  Phage‐displayed peptides as biosensor reagents , 2000, Journal of molecular recognition : JMR.

[24]  O. Thomas,et al.  High-gradient magnetic affinity separation of trypsin from porcine pancreatin. , 2002, Biotechnology and bioengineering.

[25]  J. M. Harris,et al.  Poly(Ethylene Glycol) Chemistry Biotechnical and Biomedical Applications , 1992 .

[26]  C. Mirkin,et al.  Nanoparticle-Based Bio-Bar Codes for the Ultrasensitive Detection of Proteins , 2003, Science.

[27]  G. P. Smith,et al.  Cross-linked filamentous phage as an affinity matrix. , 1998, Journal of immunological methods.

[28]  C. Mello,et al.  Chemical modification of M13 bacteriophage and its application in cancer cell imaging. , 2010, Bioconjugate chemistry.

[29]  Peng Li,et al.  Flow-enhanced nonlinear magnetophoresis for high-resolution bioseparation. , 2011, Langmuir : the ACS journal of surfaces and colloids.

[30]  Wadih Arap,et al.  Networks of gold nanoparticles and bacteriophage as biological sensors and cell-targeting agents , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[31]  Gil U. Lee,et al.  Rapid detection of dengue virus in serum using magnetic separation and fluorescence detection. , 2008, The Analyst.

[32]  Valery A. Petrenko,et al.  Landscape phage as a molecular recognition interface for detection devices , 2008, Microelectron. J..

[33]  Paul F. Barbara,et al.  Selection of peptides with semiconductor binding specificity for directed nanocrystal assembly , 2000, Nature.

[34]  W. Delano.  The PyMOL Molecular Graphics System , 2002 .

[35]  Shouheng Sun,et al.  Magnetic nanoparticles as both imaging probes and therapeutic agents. , 2010, Current topics in medicinal chemistry.

[36]  F. Szoka,et al.  Influence of multivalent nitrilotriacetic acid lipid-ligand affinity on the circulation half-life in mice of a liposome-attached His6-protein. , 2010, Bioconjugate chemistry.

[37]  Gil U. Lee,et al.  Synthesis and characterization of paramagnetic microparticles through emulsion-templated free radical polymerization. , 2006, Langmuir : the ACS journal of surfaces and colloids.

[38]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[39]  B. Mattiasson,et al.  Immobilised peptide displaying phages as affinity ligands. Purification of lactoferrin from defatted milk. , 2006, Journal of chromatography. A.

[40]  M. Russel,et al.  Filnmentous phage assembly , 1991, Molecular microbiology.

[41]  R. Tilton,et al.  Coverage-Dependent Orientation of Lysozyme Adsorbed on Silica , 2003 .

[42]  O. Thomas,et al.  High gradient magnetic separation versus expanded bed adsorption: a first principle comparison , 2001, Bioseparation.

[43]  Valery A. Petrenko,et al.  Evolution of phage display: from bioactive peptides to bioselective nanomaterials , 2008 .