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

Area of Science:

• Microbiology
• Immunology
• Genomics

Background:

• Antimicrobial resistance is driven by bacterial adaptation to selective pressures.
• Pseudomonas aeruginosa chronically infects and evolves to resist antimicrobials and evade host defenses.
• Identifying adaptive changes on the bacterial cell surface is challenging due to limited methods.

Purpose of the Study:

• To develop a high-throughput, multiplexed technology for surveying bacterial cell surfaces.
• To identify adaptive changes on the Pseudomonas aeruginosa cell surface.
• To generate novel antibodies against bacterial virulence factors.

Main Methods:

• Combining phage display with high-throughput DNA sequencing to create Phage-seq.
• Applying phage display panning to hundreds of bacterial genotypes.
• Analyzing phage display selection dynamics to capture cell surface information.

Main Results:

• Phage-seq enables high-throughput, multiplexed surveying of bacterial cell surfaces.
• The method identified key adaptive changes on the Pseudomonas aeruginosa cell surface.
• Camelid single-domain antibodies recognizing P. aeruginosa virulence factors on live cells were generated.

Conclusions:

• Phage-seq is a powerful paradigm for studying bacterial cell surfaces.
• This technology facilitates parallel identification and profiling of numerous surface features.
• The generated antibodies have potential applications in diagnostics and therapeutics for bacterial infections.