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Scalable High Throughput Selection From Phage-displayed Synthetic Antibody Libraries
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Generation of a Phage Display Chicken Single-Chain Variable Fragment Library
Hyunji Yang1, Jisu Chae1, Hyori Kim2
1Cancer Research Institute, Seoul National University College of Medicine, Seoul 03080, South Korea.
Cold Spring Harbor Protocols
|August 16, 2024
Summary
This study details creating a phage-displayed chicken antibody fragment library. This method enables antibody discovery from immunized chickens for various applications.
Area of Science:
- Immunotechnology
- Molecular Biology
- Biotechnology
Background:
- Phage-displayed antibody fragment libraries are valuable tools for antibody discovery.
- Chicken immunoglobulin genes are suitable for library construction when sequences are known.
Purpose of the Study:
- To describe a protocol for generating a phage-displayed chicken single-chain variable fragment (scFv) library.
- To facilitate antibody discovery from immunized chickens.
Main Methods:
- Amplification of rearranged heavy and light chain variable region genes.
- Linking of variable region genes via PCR to create scFv genes.
- Cloning of scFv genes into the pComb3XSS vector for phage display.
Main Results:
- Successful generation of a phage-displayed chicken scFv library.
- The protocol provides a reproducible method for library construction.
Conclusions:
- The described protocol enables the creation of chicken scFv libraries for antibody discovery.
- This method is adaptable for other species with known immunoglobulin gene sequences.
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