Bacterial cell surface characterization by phage display coupled to high-throughput sequencing

Casey N. Grun, Ruchi Jain, Maren Schniederberend, Charles B. Shoemaker, Bryce Nelson and Barbara I. Kazmierczak ()
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Casey N. Grun: Yale University School of Medicine
Ruchi Jain: Yale University School of Medicine
Maren Schniederberend: Yale University School of Medicine
Charles B. Shoemaker: Tufts Cummings School of Veterinary Medicine
Bryce Nelson: Yale University School of Medicine
Barbara I. Kazmierczak: Yale University School of Medicine

Nature Communications, 2024, vol. 15, issue 1, 1-19

Abstract: Abstract The remarkable capacity of bacteria to adapt in response to selective pressures drives antimicrobial resistance. Pseudomonas aeruginosa illustrates this point, establishing chronic infections during which it evolves to survive antimicrobials and evade host defenses. Many adaptive changes occur on the P. aeruginosa cell surface but methods to identify these are limited. Here we combine phage display with high-throughput DNA sequencing to create a high throughput, multiplexed technology for surveying bacterial cell surfaces, Phage-seq. By applying phage display panning to hundreds of bacterial genotypes and analyzing the dynamics of the phage display selection process, we capture important biological information about cell surfaces. This approach also yields camelid single-domain antibodies that recognize key P. aeruginosa virulence factors on live cells. These antibodies have numerous potential applications in diagnostics and therapeutics. We propose that Phage-seq establishes a powerful paradigm for studying the bacterial cell surface by identifying and profiling many surface features in parallel.

Date: 2024
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DOI: 10.1038/s41467-024-51912-7

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