Towards the simplification of protein synthesis: iterative solid-supported ligations with concomitant purifications

Area of Science:

• Chemical synthesis
• Biochemistry
• Organic chemistry

Background:

• Solid-phase peptide synthesis (SPPS) is a common method for creating peptides.
• Purification of synthesized peptides often requires challenging chromatographic techniques.
• Temporary peptide tagging can aid in purification processes.

Purpose of the Study:

• To develop a novel linker for temporary N-terminal peptide tagging.
• To demonstrate the utility of this linker for capture/release purification.
• To apply this strategy to the synthesis of long polypeptides.

Main Methods:

• Development of a novel linker for temporary N-terminal peptide attachment.
• Application of a self-purifying N-to-C iterative triazole ligation strategy.
• Synthesis of a 160-residue polypeptide using the developed methods.

Main Results:

• The new linker facilitates temporary tagging and capture/release purification of peptides.
• The iterative triazole ligation strategy proved highly efficient and self-purifying.
• A 160-residue polypeptide was synthesized with high purity, obviating the need for chromatography.

Conclusions:

• The developed linker and ligation strategy offer a powerful tool for peptide synthesis and purification.
• This approach significantly simplifies the production of long, pure polypeptides.
• It presents a viable alternative to traditional chromatographic purification methods.