A flow platform for degradation-free CuAAC bioconjugation
Abstract
The Cu-catalyzed azide-alkyne cycloaddition (CuAAC) reaction is a cornerstone method for the ligation of biomolecules. However, undesired Cu-mediated oxidation and Cu-contamination in bioconjugates limits biomedical utility. Here, we report a generic CuAAC flow platform for the rapid, robust, and broad-spectrum formation of discrete triazole bioconjugates. This process leverages an engineering problem to chemical advantage: solvent-mediated Cu pipe erosion generates ppm levels of Cu in situ under laminar flow conditions. This is sufficient to catalyze the CuAAC reaction of small molecule alkynes and azides, fluorophores, marketed drug molecules, peptides, DNA, and therapeutic oligonucleotides. This flow approach, not replicated in batch, operates at ambient temperature and pressure, requires short residence times, avoids oxidation of sensitive functional groups, and produces products with very low ppm Cu contamination.
Citation
Hatit , M Z C , Reichenbach , L F , Tobin , J M , Vilela , F , Burley , G A & Watson , A J B 2018 , ' A flow platform for degradation-free CuAAC bioconjugation ' , Nature Communications , vol. 9 , 4021 . https://doi.org/10.1038/s41467-018-06551-0
Publication
Nature Communications
Status
Peer reviewed
ISSN
2041-1723Type
Journal article
Description
We thank Vapourtec for the copper reactors, the EPSRC UK National Mass Spectrometry Facility at Swansea University for analyses, the University of Strathclyde for PhD studentship (M.Z.C.H.), the Leverhulme Trust for a postdoctoral fellowship (L.F.R.; grant no. RPG-2014-001), and the BBSRC for funding (grant nos. BB/R006857/1, BB/N016378/1).Collections
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