Engineered peptide barcodes for in-depth analyses of binding protein libraries.

Area of Science:

• Biotechnology
• Molecular Biology
• Biophysics

Background:

• Traditional binder generation involves laborious, individual screening of candidate molecules.
• Existing methods are time-consuming and limit the scale of binder discovery.

Purpose of the Study:

• To develop a high-throughput technology for binder selection and identification.
• To overcome limitations of current binder generation methods.

Main Methods:

• Developed NestLink, a technology using genetically encoded barcoding peptides (flycodes) for mass spectrometry detection and deep sequencing.
• Applied NestLink for simultaneous kinetic parameter ranking, deep repertoire mining, and in-cell binder identification.

Main Results:

• NestLink enables simultaneous biophysical characterization of thousands of library members.
• Successfully ranked hundreds of binder candidates by kinetic parameters.
• Identified rare binders against integral membrane proteins in cellular environments without target immobilization.

Conclusions:

• NestLink significantly accelerates binder discovery and characterization.
• The technology facilitates selection of binders with tailored characteristics.
• Opens new possibilities for in vivo and in situ binder selection.