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A Protocol for Phage Display and Affinity Selection Using Recombinant Protein Baits
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Identification of immunogenic commensal antigens using phage display
Sheenam Verma1, Samantha Kimmel1,2, Oliver J Harrison3,4
1Center for Fundamental Immunology, Benaroya Research Institute, Seattle, WA, USA.
Nature Protocols
|June 17, 2025
Summary
This study introduces a novel bacteriophage display protocol to identify microbial antigens that trigger immune responses. This method aids in understanding host-microbe interactions and developing new diagnostics or therapeutics.
Area of Science:
- Microbiology
- Immunology
- Molecular Biology
Background:
- Humoral immunity is crucial for host-microbe interactions and defense against pathogens.
- Identifying specific antigens that elicit adaptive immune responses remains a challenge.
Purpose of the Study:
- To present a protocol for identifying immunogenic protein antigens from microbes using bacteriophage display.
- To enable the study of host-microbe dialogue and immune responses.
Main Methods:
- Generation of an M13-phagemid library and an ORFeome phage library.
- Affinity-based immunoprecipitation and antigen validation using ELISA, ELISPOT, and B cell tetramers.
- Identification of antigens via Sanger sequencing of monoclonal phages.
Main Results:
- The protocol successfully identifies antigens eliciting IgA and IgG responses.
- It can utilize circulating or intestinal antibodies for antigen discovery.
- Provides protein domain-level resolution of epitope-bearing regions.
Conclusions:
- This bacteriophage display protocol offers an efficient method for antigen identification and validation.
- It can be performed by trained molecular biologists within weeks.
- Facilitates deeper understanding of humoral immunity in host-microbe interactions.
