Synthetic antibodies designed on natural sequence landscapes

Area of Science:

• Immunology
• Synthetic Biology
• Biotechnology

Background:

• Antibody libraries are crucial for drug discovery and research.
• Existing synthetic antibody libraries often lack the diversity and fidelity of natural repertoires.

Purpose of the Study:

• To develop a novel method for generating large-scale synthetic antibody libraries.
• To create libraries that accurately mimic natural antibody diversity while enabling access to novel sequences.
• To demonstrate the functional utility of the generated library for antibody isolation.

Main Methods:

• High-throughput immune repertoire analysis to inform library design.
• Novel gene synthesis technology for precise control of amino acid composition and frequency.
• Construction and high-throughput pyrosequencing of a synthetic antibody library with 3.6×10^10 transformants.
• Fab expression analysis and genetic diversity characterization.

Main Results:

• The synthetic library demonstrated superior Fab expression (>93% correctly folded heavy and light chains) compared to existing methods.
• High genetic diversity was achieved, with 95% of sequenced clones appearing only once.
• The library successfully isolated antibodies with low and sub-nanomolar affinities against various targets.

Conclusions:

• The developed method successfully generates diverse and functional synthetic antibody libraries.
• This approach provides a powerful tool for antibody discovery and engineering, bridging natural repertoire diversity with synthetic accessibility.
• The strategy is validated by the isolation of high-affinity antibodies against a wide range of targets.