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

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

Abstract

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 

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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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