Bacteria-based in vivo peptide library screening using biopanning approach

Area of Science:

• Biochemistry
• Molecular Biology
• Drug Discovery

Background:

• Traditional library screening for therapeutic agents typically occurs in vitro using purified components.
• In vitro identified agents often lack efficacy in cellular environments.
• A need exists for screening systems that identify biologically active molecules within a cellular context.

Purpose of the Study:

• To develop and validate an in vivo peptide library screening system.
• To identify peptides that inhibit protein-protein interactions within living cells.
• To provide a faster and more effective method for discovering therapeutic peptides.

Main Methods:

• Developed an in vivo screening system utilizing bacteriophage lambda's cI repressor reconstitution in bacterial cells.
• Employed a high-density expression peptide library for screening.
• Utilized a multiple-round biopanning approach for peptide enrichment and amplification.

Main Results:

• Successfully established a functional in vivo screening platform within bacterial cells.
• Demonstrated the system's capability to identify peptides that interfere with protein assembly.
• The biopanning approach enhanced the identification of positive peptide hits.

Conclusions:

• The developed in vivo system offers a rapid and efficient method for discovering biologically active peptides.
• This approach overcomes the limitations of in vitro screening for identifying cell-active therapeutic agents.
• The system is suitable for identifying inhibitors of target protein interactions and assembly.