Facile Preparation of PNA-Peptide Conjugates with a Polar Maleimide-Thioether Linkage
Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review
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.
Original language | English |
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Title of host publication | Peptide Nucleic Acids : Methods and Protocols |
Editors | Peter E. Nielsen |
Number of pages | 22 |
Publisher | Humana Press |
Publication date | 2020 |
Pages | 97-118 |
ISBN (Print) | 978-1-0716-0242-3, 978-1-0716-0245-4 |
DOIs | |
Publication status | Published - 2020 |
Series | Methods in molecular biology (Clifton, N.J.) |
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Volume | 2105 |
ISSN | 1064-3745 |
ID: 269722976