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Solid Phase Synthesis of a Functionalized Bis-Peptide Using "Safety Catch" Methodology
Published on: May 15, 2012
Solid-phase peptide synthesis: from standard procedures to the synthesis of difficult sequences
Irene Coin1, Michael Beyermann, Michael Bienert
1Leibniz-Institut für Molekulare Pharmakologie, Department of Peptide Chemistry and Biochemistry, Robert-Rössle-Strasse 10, Berlin 13125, Germany.
Nature Protocols
|December 15, 2007
Summary
This study details a robust solid-phase peptide synthesis (SPPS) protocol using the Fmoc/tBu strategy for efficient peptide production. Special techniques are introduced to overcome challenges in synthesizing difficult peptide sequences, improving success rates in peptide chemisynthesis.
Area of Science:
- Organic Chemistry
- Biochemistry
- Synthetic Chemistry
Background:
- Solid-phase peptide synthesis (SPPS) is a cornerstone of peptide production.
- The Fmoc/tBu strategy is widely adopted for its efficiency and orthogonality.
- Challenges remain in synthesizing complex or aggregation-prone peptide sequences.
Purpose of the Study:
- To present a refined SPPS protocol based on the Fmoc/tBu strategy.
- To demonstrate successful synthesis of standard and complex peptides.
- To introduce methods for overcoming difficulties in peptide chemisynthesis.
Main Methods:
- Utilized the Fmoc/tBu strategy with aminium-derived coupling reagents.
- Employed PEG-modified polystyrene resins for solid-phase synthesis.
- Applied specialized techniques, including depsipeptide and pseudoproline units, for difficult sequences.
Main Results:
- Successfully synthesized the 41-mer peptide corticotropin-releasing factor (CRF) in approximately 80 working hours.
- Produced over 400 CRF analogs and numerous other peptides.
- Achieved synthesis of an extremely difficult sequence (Asn(15) analog of WW domain FBP28) using specialized methods, which failed with the standard protocol.
Conclusions:
- The described SPPS protocol is effective for standard peptide synthesis, including CRF.
- Specialized techniques are crucial for overcoming aggregation and synthesizing challenging peptide sequences.
- This approach enhances the scope and success rate of peptide chemisynthesis.
