An Integrated Workflow for Phage Selection, Amplification, and Surface Modification

Bacteriophages, characterized by strong host specificity and high environmental compatibility, are widely recognized for their immense potential in microbial ecological regulation. They can be utilized for foodborne pathogenic bacteria control and drug-resistant bacterial infections treatment. To address the limitations of traditional phage screening methods when isolating target phages from complex environmental samples, a standardized and efficient technical protocol can be established by integrating optimized techniques, including the double-layer plate method, liquid enrichment combined with centrifugation, and supermicro buffer-mediated phage stability maintenance. In this chapter, we describe protocols for the preparation of key reagents, the implementation of a complete phage screening workflow, phage amplification via liquid/plate/OD₂₆₀ optical density methods, and chemical coupling-based phage surface modification to achieve functionalization, along with critical notes to ensure reproducibility and reliability of the experimental process.