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Facile Preparation of PNA-Peptide Conjugates with a Polar Maleimide-Thioether Linkage

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Peptide Nucleic Acids

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2105))

Abstract

Conjugation of a delivery peptide containing a thiol functionality (e.g., a cysteine residue) with a PNA oligomer displaying a single unprotected aliphatic primary amine (e.g., the N-terminus or a C-terminal lysine residue) can be achieved via a one-pot modification with a bisfunctional maleimide linker also displaying a reactive N-hydroxysuccinimidyl ester group (e.g., Mal-PEG2-OSu). Here, an optimized protocol with respect to ratios between the reactants as well as recommended reaction times is presented. Formation and conversion of the maleimide-PNA intermediate was followed by analytical HPLC as exemplified by its conjugation to (KFF)3K-Cys-NH2. In addition, the reaction time required for direct conversion of a preformed Mal-(CH2)2-(C=O)-PNA oligomer in the presence of a slight excess of thiol-modified peptide (with a varying degree of sterical hindrance: HS-(CH2)2-CONH-(KFF)3K-NH2, (KFF)3K-hCys-NH2 and (KFF)3K-Cys-NH2) is provided.

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Acknowledgments

This research was supported by the Department of Drug Design and Pharmacology and Center for Peptide-Based Antibiotics (Cepan).

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Authors and Affiliations

  1. Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark

    Anna Mette Hansen, Ashif Yasin Shaikh & Henrik Franzyk

Authors
  1. Anna Mette Hansen
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  2. Ashif Yasin Shaikh
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  3. Henrik Franzyk
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Corresponding author

Correspondence to Henrik Franzyk .

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Editors and Affiliations

  1. Department of Cellular and Molecular Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark

    Peter E. Nielsen

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Hansen, A.M., Shaikh, A.Y., Franzyk, H. (2020). Facile Preparation of PNA-Peptide Conjugates with a Polar Maleimide-Thioether Linkage. In: Nielsen, P. (eds) Peptide Nucleic Acids. Methods in Molecular Biology, vol 2105. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0243-0_6

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  • DOI: https://doi.org/10.1007/978-1-0716-0243-0_6

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0242-3

  • Online ISBN: 978-1-0716-0243-0

  • eBook Packages: Springer Protocols

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