Abstract
The increasing development of bacterial resistance to traditional antibiotics has reached alarming levels, thus there is an urgent need to develop new antimicrobial agents. To be effective, these new antimicrobials should possess novel modes of action and/or different cellular targets compared with existing antibiotics. Bacteriophages (phages) have been used for over a century as tools for the treatment of bacterial infections, for nearly half a century as tools in genetic research, for about two decades as tools for the discovery of specific target-binding proteins and peptides, and for almost a decade as tools for vaccine development. We describe a new application in the area of antibacterial nanomedicines where filamentous phages can be formulated as targeted drug-delivery vehicles of nanometric dimensions (phage nanomedicines) and used for therapeutic purposes. This protocol involves both genetic and chemical engineering of these phages. The genetic engineering of the phage coat, which results in the display of a target-specificity-conferring peptide or protein on the phage coat, can be used to design the drug-release mechanism and is not described herein. However, the methods used to chemically conjugate cytotoxic drugs at high density on the phage coat are described. Further, assays to measure the drug load on the surface of the phage and the potency of the system in the inhibition of growth of target cells as well as assessment of the therapeutic potential of the phages in a mouse disease model are discussed.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Yoshikawa, T. T. (2002) Antimicrobial resistance and aging: beginning of the end of the antibiotic era? J. Am. Geriatr. Soc. 50, S226–S229.
Forget, E. J. and Menzies, D. (2006) Adverse reactions to first-line antituberculosis drugs. Expert Opin. Drug Saf. 5, 231–249.
Yacoby, I., Shamis, M., Bar, H., Shabat, D., and Benhar, I. (2006) Targeting antibacterial agents by using drug-carrying filamentous bacteriophages. Antimicrob. Agents Chemother. 50, 2087–2097.
Yacoby, I., Bar, H., and Benhar, I. (2007) Targeted drug-carrying bacteriophages as antibacterial nanomedicines. Antimicrob. Agents Chemother. 51, 2156–2163.
Smith, G. P. (1985) Filamentous fusion phage: novel expression vectors that display cloned antigens on the virion surface. Science 228, 1315–1317.
Beckett, D., Kovaleva, E., and Schatz, P. J. (1999) A minimal peptide substrate in biotin holoenzyme synthetase-catalyzed biotinylation. Protein Sci. 8, 921–929.
Scholle, M. D., Kriplani, U., Pabon, A., Sishtla, K., Glucksman, M. J., and Kay, B. K. (2006) Mapping protease substrates by using a biotinylated phage substrate library. Chembiochem 7, 834–838.
Zou, J., Dickerson, M. T., Owen, N. K., Landon, L. A., and Deutscher, S. L. (2004) Biodistribution of filamentous phage peptide libraries in mice. Mol. Biol. Rep. 31, 121–129.
Molenaar, T. J., Michon, I., de Haas, S. A., van Berkel, T. J., Kuiper, J., and Biessen, E. A. (2002) Uptake and processing of modified bacteriophage M13 in mice: implications for phage display. Virology 293, 182–191.
Benhar, I. (2001) Biotechnological applications of phage and cell display. Biotechnol. Adv. 19, 1–33.
Cortese, R., Felici, F., Galfre, G., Luzzago, A., Monaci, P., and Nicosia, A. (1994) Epitope discovery using peptide libraries displayed on phage. Trends Biotechnol. 12, 262–267.
Hoogenboom, H. R., de Bruine, A. P., Hufton, S. E., Hoet, R. M., Arends, J. W., and Roovers, R. C. (1998) Antibody phage display technology and its applications. Immunotechnology 4, 1–20.
Sidhu, S. S., Weiss, G. A., and Wells, J. A. (2000) High copy display of large proteins on phage for functional selections. J. Mol. Biol. 296, 487–495.
Bar, H., Yacoby, I., and Benhar, I. (2008) Killing cancer cells by targeted drug-carrying phage nanomedicines. BMC Biotechnol. 8, 37.
Berkowitz, S. A. and Day, L. A. (1976) Mass, length, composition and structure of the filamentous bacterial virus fd. J. Mol. Biol. 102, 531–547.
Enshell-Seijffers, D., Smelyanski, L., and Gershoni, J. M. (2001) The rational design of a “type 88” genetically stable peptide display vector in the filamentous bacteriophage fd. Nucleic Acids Res. 29, E50–E60.
Nilsson, B., Moks, T., Jansson, B., Abrahmsen, L., Elmblad, A., Holmgren, E., et al. (1987) A synthetic IgG-binding domain based on staphylococcal protein A. Protein Eng. 1, 107–113.
Nilsson, J., Larsson, M., Stahl, S., Nygren, P. A., and Uhlen, M. (1996) Multiple affinity domains for the detection, purification and immobilization of recombinant proteins. J. Mol. Recognit. 9, 585–594.
Enshell-Seijffers, D. and Gershoni, J. M. (2002) Phage display selection and analysis of Ab-binding epitopes. in: Current Protocols in Immunology (Coligan, J. E., Bierer, B. E., Margulies, D. H., Shevach, E. M., and Strober, W., eds) pp. 9.8.1-9.8.27. John Wiley & Sons, Inc., USA.
Acknowledgments
Studies of targeted drug-carrying phage nanomedicines at the author’s laboratory received a grant from the Israel Public Committee for Allocation of Estate Funds, Ministry of Justice, Israel and by the Israel Cancer Association.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Vaks, L., Benhar, I. (2011). Antibacterial Application of Engineered Bacteriophage Nanomedicines: Antibody-Targeted, Chloramphenicol Prodrug Loaded Bacteriophages for Inhibiting the Growth of Staphylococcus aureus Bacteria. In: Hurst, S. (eds) Biomedical Nanotechnology. Methods in Molecular Biology, vol 726. Humana Press. https://doi.org/10.1007/978-1-61779-052-2_13
Download citation
DOI: https://doi.org/10.1007/978-1-61779-052-2_13
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-051-5
Online ISBN: 978-1-61779-052-2
eBook Packages: Springer Protocols