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
Standard
Facile Preparation of PNA-Peptide Conjugates with a Polar Maleimide-Thioether Linkage. / Hansen, Anna Mette; Shaikh, Ashif Yasin; Franzyk, Henrik.
Peptide Nucleic Acids: Methods and Protocols. ed. / Peter E. Nielsen. Humana Press, 2020. p. 97-118 (Methods in molecular biology (Clifton, N.J.), Vol. 2105).Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review
Harvard
APA
Vancouver
Author
Bibtex
}
RIS
TY - CHAP
T1 - Facile Preparation of PNA-Peptide Conjugates with a Polar Maleimide-Thioether Linkage
AU - Hansen, Anna Mette
AU - Shaikh, Ashif Yasin
AU - Franzyk, Henrik
PY - 2020
Y1 - 2020
N2 - 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.
AB - 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.
U2 - 10.1007/978-1-0716-0243-0_6
DO - 10.1007/978-1-0716-0243-0_6
M3 - Book chapter
C2 - 32088866
SN - 978-1-0716-0242-3
SN - 978-1-0716-0245-4
T3 - Methods in molecular biology (Clifton, N.J.)
SP - 97
EP - 118
BT - Peptide Nucleic Acids
A2 - null, Peter E. Nielsen
PB - Humana Press
ER -
ID: 269722976