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Deep sequencing in library selection projects: what insight does it bring?
J Glanville1, S D'Angelo2, T A Khan3
1Program in Computational and Systems Immunology, Institute for Immunity, Transplantation and Infection, Stanford University School of Medicine, Stanford, CA, USA.
Current Opinion in Structural Biology
|October 10, 2015
Summary
High throughput sequencing revolutionizes antibody discovery and engineering. This technology enables deep insights into antibody libraries and selections, guiding improved antibody design and development.
Area of Science:
- Biotechnology
- Immunology
- Bioinformatics
Background:
- Antibody discovery and engineering are critical in therapeutics and diagnostics.
- Traditional methods face limitations in scale and precision.
- Advancements in sequencing technologies offer new possibilities.
Purpose of the Study:
- To review the impact of high throughput sequencing on antibody discovery and engineering.
- To outline key considerations for successful implementation of these technologies.
- To discuss current challenges and future directions.
Main Methods:
- High throughput sequencing for library design and selection analysis.
- Polymerase Chain Reaction (PCR) reagent design.
- Selection of appropriate sequencing platforms.
- Computational tools and statistical analysis for data interpretation.
Main Results:
- Enables creation of large, high-quality antibody libraries with billions of unique molecules.
- Facilitates quantitative tracing of enriched clones during selection.
- Provides position-specific insights into amino acid variations under selection pressure.
- Allows for error removal, diversity estimation, and identification of selection signatures.
Conclusions:
- High throughput sequencing significantly enhances antibody discovery and engineering processes.
- Successful application requires careful attention to PCR reagents, sequencing platforms, and computational analysis.
- Addressing remaining challenges will accelerate widespread adoption and further innovation.
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