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Updated: May 8, 2025
Bacterial Inner-membrane Display for Screening a Library of Antibody Fragments
Published on: October 15, 2016
Phage Display Isolation of VHH Antibodies Targeting Capsular Polysaccharides of Intratumor Bacteria
Qun Ji1,2, Oren Moscovitz3
1Department of Biomolecular Systems, Max Planck Institute of Colloids and Interfaces, Potsdam, Germany.
Abstract:
Phage display, a well-established and powerful tool, has been used to identify and isolate antibody fragments against a range of diverse cancer-related antigen types. Nonetheless, the availability of specific antibodies and antibody fragments against carbohydrate antigens remains low and insufficient. Bacteria found within cancer and immune cells affect tumorigenesis, progression, and shaping of the immunosuppressive microenvironment. Yet, their identification primarily relies on nucleic acid-based sequencing techniques at the species level, leaving a significant gap in molecular tools that distinguish bacterial strains and serotypes. This chapter provides a comprehensive protocol for phage display-based VHH antibodies identification against the capsular polysaccharides that define bacterial serotypes based on their composition and structure. We aim to illuminate a path for developing antibody fragments that will enable the targeting of specific intracellular bacterial strains and serotypes in tumor and immune cells, thereby unraveling their role and contribution to cancer biology.
Insights
This study introduces a phage display method to create VHH antibodies targeting bacterial capsular polysaccharides. This advances the identification of specific bacterial strains and serotypes within cancer cells, aiding cancer research.
Area of Science:
- Oncology
- Microbiology
- Immunology
- Biotechnology
Background:
- Phage display is effective for identifying antibodies against cancer antigens, but lacks specificity for carbohydrate antigens.
- Intracellular bacteria influence cancer development and the tumor microenvironment, yet are typically identified only at the species level.
- Current methods fail to distinguish between bacterial strains and serotypes, limiting understanding of their role in cancer.
Purpose of the Study:
- To develop a protocol for identifying VHH antibodies against bacterial capsular polysaccharides using phage display.
- To enable the development of molecular tools for precise targeting of bacterial strains and serotypes within tumor and immune cells.
- To investigate the role of specific intracellular bacteria in cancer biology and the immunosuppressive tumor microenvironment.
Main Methods:
- Phage display technology was employed to select and identify VHH antibody fragments.
- The protocol focuses on targeting capsular polysaccharides, which are key components of bacterial serotypes.
- The VHH antibodies were generated against specific bacterial serotype-defining capsular polysaccharides.
Main Results:
- A comprehensive protocol for generating VHH antibodies against bacterial capsular polysaccharides via phage display is presented.
- The developed VHH antibodies show potential for distinguishing specific bacterial strains and serotypes.
- This approach facilitates the creation of tools to target intracellular bacteria within the tumor microenvironment.
Conclusions:
- Phage display offers a viable strategy for developing specific VHH antibodies against bacterial carbohydrate antigens.
- This method addresses the current gap in molecular tools for identifying and targeting specific bacterial strains and serotypes in cancer.
- The VHH antibodies developed can aid in unraveling the contribution of intracellular bacteria to cancer progression and immune evasion.
