High-throughput protein binder discovery by rapid in vivo selection

Area of Science:

• Biotechnology
• Molecular Biology
• Protein Engineering

Background:

• Protein binders are crucial for research, diagnostics, and therapeutics.
• Existing binder discovery methods are slow, costly, and have high failure rates, especially for challenging targets.

Purpose of the Study:

• To establish a novel, high-throughput *in vivo* platform for rapid and efficient discovery of protein binders.
• To overcome limitations of current binder discovery techniques.

Main Methods:

• Developed Phage-Assisted Non-Continuous Selection of Protein Binders (PANCS-binders).
• Utilized M13 phage and proximity-dependent split RNA polymerase biosensors for target binding linkage.
• Screened billion-plus member protein variant libraries against multiple therapeutic targets.

Main Results:

• Successfully screened over 10^11 protein-protein interaction pairs in two days.
• Generated large, high-quality datasets and identified hundreds of novel protein binders.
• Demonstrated that identified binders can be affinity matured or directly used in mammalian cells.

Conclusions:

• PANCS-binders significantly accelerates and simplifies the protein binder discovery process.
• This platform has the potential to democratize binder discovery and advance proteome-targeting biotechnologies.