Journal
CHEMBIOCHEM
Volume 17, Issue 9, Pages 866-875Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/cbic.201500582
Keywords
biocompatibility; click chemistry; coppercatalyzed azide-alkyne cycloaddition; glycoconjugates; glycolabeling efficiency; strain-promoted azide-alkyne cycloaddition
Funding
- DAAD grant, Kooperation in Pharmazeutische Wissenschaften und Lehre [57058983]
- European project MANAQA, Magnetic Nano Actuators for Quantitative Analysis [296679]
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Bio-orthogonal copper (I)-catalyzed azide-alkyne cycloaddition (CuAAC) has been widely used to modify azide-or alkyne-bearing monosaccharides on metabolic glyco-engineered mammalian cells. Here, we present a systematic study to elucidate the design space for the cytotoxic effects of the copper catalyst on NIH 3T3 fibroblasts and on HEK 293-F cells. Monitoring membrane integrity by flow cytometry and RT-PCR analysis with apoptotic and anti-apoptotic markers elucidated the general feasibility of CuAAC, with exposure time of the CuAAC reaction mixture having the major influence on biocompatibility. A high labeling efficiency of HEK293-F cells with a fluorescent alkyne dye was rapidly achieved by CuAAC in comparison to copper free strain-promoted azide-alkyne cycloaddition (SPAAC). The study details effective and biocompatible conditions for CuAAC-based modification of glyco-engineered cells in comparison to its copper free alternative.
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