Isolation of Competitive Phage Display-Modified Bacteriophage T4 with Affinity Chromatography

  • Krystyna DąbrowskaEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1898)


Phage recovery from various solutions, including physiological samples, as well as phage purification from crude lysates often requires a specific isolation method. Here, we demonstrate that T4-like phages can be efficiently isolated by affinity chromatography. This approach employs specific affinity tags (GST (glutathione S-transferase) or His-tag) that allow for the isolation of the phage. These affinity tags are exposed on the phage head using phage display. By combining competitive phage display and affinity chromatography, wild-type phages can be specifically recovered from mixtures with other phage/s, from solutions of very low phage concentration, or purified from crude phage lysates.

Key words

Phage purification Affinity chromatography Competitive phage display T4 bacteriophage Hoc protein Escherichia coli 


  1. 1.
    Chibani Azaïez SR, Fliss I, Simard RE et al (1998) Monoclonal antibodies raised against native major capsid proteins of lactococcal c2-like bacteriophages. Appl Environ Microbiol 64:4255–4259PubMedPubMedCentralGoogle Scholar
  2. 2.
    Shelton CB, Crosslin DR, Casey JL et al (2000) Discovery, purification, and characterization of a temperate transducing bacteriophage for Bordetella avium. J Bacteriol 182:6130–6136CrossRefGoogle Scholar
  3. 3.
    McLaughlin MR, King RA (2008) Characterization of Salmonella bacteriophages isolated from swine lagoon effluent. Curr Microbiol 56:208–213CrossRefGoogle Scholar
  4. 4.
    Boratyński J, Syper D, Weber-Dabrowska B et al (2004) Preparation of endotoxin-free bacteriophages. Cell Mol Biol Lett 9:253–259PubMedGoogle Scholar
  5. 5.
    Brorson K, Shen H, Lute S et al (2008) Characterization and purification of bacteriophages using chromatofocusing. J Chromatogr A 1207:110–121CrossRefGoogle Scholar
  6. 6.
    Kramberger P, Honour RC, Herman RE et al (2010) Purification of the Staphylococcus aureus bacteriophages VDX-10 on methacrylate monoliths. J Virol Methods 166:60–64CrossRefGoogle Scholar
  7. 7.
    Adriaenssens EM, Lehman SM, Vandersteegen K et al (2012) CIM(®) monolithic anion-exchange chromatography as a useful alternative to CsCl gradient purification of bacteriophage particles. Virology 434:265–270CrossRefGoogle Scholar
  8. 8.
    Oksanen HM, Domanska A, Bamford DH (2012) Monolithic ion exchange chromatographic methods for virus purification. Virology 434:271–277CrossRefGoogle Scholar
  9. 9.
    Ceglarek I, Piotrowicz A, Lecion D et al (2013) A novel approach for separating bacteriophages from other bacteriophages using affinity chromatography and phage display. Sci Rep 3:3220CrossRefGoogle Scholar
  10. 10.
    Sambrook J, Russell DW (eds) (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory, New YorkGoogle Scholar
  11. 11.
    Adams MH (1956) Bacteriophages. Inter Science Publication, New YorkGoogle Scholar
  12. 12.
    Ren Z, Black LW (1998) Phage T4 SOC and HOC display of biologically active, full-length proteins on the viral capsid. Gene 215:439–444CrossRefGoogle Scholar
  13. 13.
    Shivachandra SB, Li Q, Peachman KK et al (2007) Multicomponent anthrax toxin display and delivery using bacteriophage T4. Vaccine 25:1225–1235CrossRefGoogle Scholar
  14. 14.
    Jiang J, Abu-Shilbayeh L, Rao VB (1997) Display of a PorA peptide from Neisseria meningitidis on the bacteriophage T4 capsid surface. Infect Immun 65:4770–4777PubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Bacteriophage LaboratoryInstitute of Immunology and Experimental Therapy, Polish Academy of SciencesWrocławPoland

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