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Biocompatible Azide–Alkyne “Click” Reactions for Surface Decoration of Glyco‐Engineered Cells
Author(s) -
Gutmann Marcus,
Memmel Elisabeth,
Braun Alexandra C.,
Seibel Jürgen,
Meinel Lorenz,
Lühmann Tessa
Publication year - 2016
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201500582
Subject(s) - cycloaddition , alkyne , azide , click chemistry , biocompatibility , chemistry , surface modification , combinatorial chemistry , biocompatible material , bioorthogonal chemistry , flow cytometry , biophysics , biochemistry , catalysis , organic chemistry , biomedical engineering , microbiology and biotechnology , biology , medicine
Abstract 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 HEK 293‐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.