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Scalable High Throughput Selection From Phage-displayed Synthetic Antibody Libraries
Published on: January 17, 2015
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A High-Throughput Single-Clone Phage Fluorescence Microwell Immunoassay and Laser-Driven Clonal Retrieval System
Seohee Chang1, Soohyun Kim2,3, Jerome Han2,4
1Department of Electrical and Computer Engineering, Seoul National University, Seoul 08826, Korea.
Biomolecules
|April 3, 2020
Summary
We developed a high-throughput screening system to improve antibody discovery using phage display. This method screens more clones, yielding diverse antigen binders and potentially reducing biopanning rounds.
Area of Science:
- Biotechnology
- Molecular Biology
- Immunology
Background:
- Phage display is a key technology for therapeutic antibody selection, but faces limitations like asymmetrical amplification.
- These limitations can restrict the diversity and number of antigen binders obtained from libraries.
Purpose of the Study:
- To develop a high-throughput single-clonal screening system for antibody discovery.
- To overcome the limitations of traditional phage display biopanning.
- To increase the diversity of antigen-reactive antibody fragments.
Main Methods:
- Developed a system combining fluorescence immunoassays with laser-driven clonal DNA retrieval using microchip technology.
- Screened over 70,000 clones (~14% of library complexity) from a single-chain variable fragment (scFv) phage library.
- Focused on libraries with random mutations at five amino acid residues.
Main Results:
- Identified 78 antigen-reactive scFv sequences with mutations limited to the randomized residues.
- Demonstrated screening of a significant portion of library complexity in a single run.
- Achieved a higher yield of specific binders compared to conventional methods.
Conclusions:
- The developed system significantly enhances the efficiency of antibody screening.
- It can reduce or eliminate the need for multiple biopanning rounds.
- Offers a pathway to access a more diverse repertoire of antibody clones.
