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Rapid solid-phase synthesis of a calmodulin-binding peptide using controlled microwave irradiation

Nature Protocols volume 2, pages 2222–2227 (2007)Cite this article

Abstract

A rapid and efficient microwave-assisted solid-phase synthesis method for the preparation of a nonapeptide using conventional Fmoc/But orthogonal protection strategy is described. In this protocol, the coupling steps are performed within 5 min at 60 °C and the Fmoc-deprotection steps are completed within 3 min at 60 °C using a dedicated single-mode microwave peptide synthesizer utilizing temperature-controlled conditions. It is demonstrated that the model nonapeptide (containing the calmodulin-binding octapeptide sequence) is synthesized in a shorter time (∼3.5 h) and with high purity (>95%) under microwave irradiation conditions in comparison with a reference peptide that is obtained by standard methods at room temperature (within 11 h).

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Figure 1
Figure 2
Figure 3: Temperature profile for a microwave-assisted peptide coupling step (Fmoc-amino acid, DIC, HOBt, NMP and resin; ca. 1 ml total volume) heating to 60 °C (10 W maximum magnetron power) for 5 min in a single-mode microwave peptide synthesizer.
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Acknowledgements

We thank piCHEM R&D for access to the HPLC, lyophilizer and MALDI-TOF MS.

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  1. Christian Doppler Laboratory for Microwave Chemistry (CDLMC) and Institute of Chemistry, Karl-Franzens-University Graz, Heinrichstrasse 28, Graz, A-8010, Austria

    Bernadett Bacsa & C Oliver Kappe

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Correspondence to C Oliver Kappe.

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Bacsa, B., Kappe, C. Rapid solid-phase synthesis of a calmodulin-binding peptide using controlled microwave irradiation. Nat Protoc 2, 2222–2227 (2007). https://doi.org/10.1038/nprot.2007.300

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