Immobilized Thiol Catalysts for Native Chemical Ligation in Flow
5 affiliations
- 1Novo Nordisk A/S , , Smørmosevej 17-19 , ,
- 2Bachem AG , Hauptstrasse 144 , ,
- 3Novo Nordisk Pharmatech A/S , Københavnsvej 216 , ,
- 4Novo Nordisk A/S , , ,
- 5University of Copenhagen , , Jagtvej 160 , ,
Abstract
Native chemical ligation (NCL) is a well-established research tool used to access long and complex peptides that cannot be efficiently synthesized by other means such as linear solid-phase peptide synthesis (SPPS). However, its broader application beyond the research laboratory scale has been limited by low-volume yields, slow reaction kinetics, and challenges associated with homogeneous thiol catalysts, as these catalysts can be incompatible with desulfurization methods and difficult to remove in subsequent chromatographic purification. In this study, we report a continuous-flow NCL platform enabled by the development of a novel immobilized version of the aromatic thiol catalyst 4-mercaptophenylacetic acid (MPAA). This strategy employs shelf-stable peptide hydrazides that are activated in situ to acyl pyrazoles and subsequently converted to reactive thioesters in flow by the immobilized catalyst, thereby facilitating ligation with cysteine-containing peptides. The platform is directly compatible with inline desulfurization and supports high-throughput synthesis of complex, structurally diverse, and therapeutically relevant peptides up to 84 amino acids in length. This work establishes immobilized aryl thiols as viable catalysts for NCL in continuous flow, identifies involved key challenges, and provides a foundation for its prospects toward industrial peptide manufacturing.
Versions
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BibTeX
@article{egelund2026immobilized,
title = {{Immobilized Thiol Catalysts for Native Chemical Ligation in Flow}},
author = {Peter H. G. Egelund and Jonathan Pfeuffer-Rooschüz and Sandip Jadhav and Iacopo Galleano and François Crestey and Martin Lund and Felix Wojcik and Ralph Schoenleber and Christian A. Olsen and Henrik Johansson Castro and Carolin Lechner and Daniel Sejer Pedersen},
journal = {JACS Au},
year = {2026},
month = oct,
doi = {10.1021/jacsau.6c01030},
url = {https://pubs.acs.org/jaaucr/article/doi/10.1021/jacsau.6c01030/5453236/Immobilized-Thiol-Catalysts-for-Native-Chemical},
note = {CC BY-NC-ND 4.0},
}
RIS
TY - JOUR
TI - Immobilized Thiol Catalysts for Native Chemical Ligation in Flow
AU - Peter H. G. Egelund
AU - Jonathan Pfeuffer-Rooschüz
AU - Sandip Jadhav
AU - Iacopo Galleano
AU - François Crestey
AU - Martin Lund
AU - Felix Wojcik
AU - Ralph Schoenleber
AU - Christian A. Olsen
AU - Henrik Johansson Castro
AU - Carolin Lechner
AU - Daniel Sejer Pedersen
PY - 2026
DA - 2026/10/17
JO - JACS Au
DO - 10.1021/jacsau.6c01030
UR - https://pubs.acs.org/jaaucr/article/doi/10.1021/jacsau.6c01030/5453236/Immobilized-Thiol-Catalysts-for-Native-Chemical
AB - Native chemical ligation (NCL) is a well-established research tool used to access long and complex peptides that cannot be efficiently synthesized by other means such as linear solid-phase peptide synthesis (SPPS). However, its broader application beyond the research laboratory scale has been limited by low-volume yields, slow reaction kinetics, and challenges associated with homogeneous thiol catalysts, as these catalysts can be incompatible with desulfurization methods and difficult to remove in subsequent chromatographic purification. In this study, we report a continuous-flow NCL platform enabled by the development of a novel immobilized version of the aromatic thiol catalyst 4-mercaptophenylacetic acid (MPAA). This strategy employs shelf-stable peptide hydrazides that are activated in situ to acyl pyrazoles and subsequently converted to reactive thioesters in flow by the immobilized catalyst, thereby facilitating ligation with cysteine-containing peptides. The platform is directly compatible with inline desulfurization and supports high-throughput synthesis of complex, structurally diverse, and therapeutically relevant peptides up to 84 amino acids in length. This work establishes immobilized aryl thiols as viable catalysts for NCL in continuous flow, identifies involved key challenges, and provides a foundation for its prospects toward industrial peptide manufacturing.
SN - 2691-3704
ER -