title: Biocompatible Azide-Alkyne "Click" Reactions for Surface Decoration of Glyco-Engineered Cells.
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.
authors.affiliation: Institute of Pharmacy and Food Chemistry, University of Würzburg, Am Hubland, 97074, Würzburg, Germany.
authors.forename: Marcus
authors.initials: M
authors.lastname: Gutmann
authors.affiliation: Institute of Organic Chemistry, University of Würzburg, Am Hubland, 97074, Würzburg, Germany.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Memmel
authors.affiliation: Institute of Pharmacy and Food Chemistry, University of Würzburg, Am Hubland, 97074, Würzburg, Germany.
authors.forename: Alexandra C
authors.initials: AC
authors.lastname: Braun
authors.affiliation: Institute of Organic Chemistry, University of Würzburg, Am Hubland, 97074, Würzburg, Germany.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Seibel
authors.affiliation: Institute of Pharmacy and Food Chemistry, University of Würzburg, Am Hubland, 97074, Würzburg, Germany.
authors.forename: Lorenz
authors.initials: L
authors.lastname: Meinel
authors.affiliation: Institute of Pharmacy and Food Chemistry, University of Würzburg, Am Hubland, 97074, Würzburg, Germany. tessa.luehmann@uni-wuerzburg.de.
authors.forename: Tessa
authors.initials: T
authors.lastname: Lühmann
mesh_terms: D000480:Alkynes / Q000737:chemistry*; D000818:Animals; D051017:Apoptosis Regulatory Proteins / Q000235:genetics / Q000378:metabolism; D001386:Azides / Q000737:chemistry*; D001672:Biocompatible Materials / Q000737:chemistry* / Q000633:toxicity; D002384:Catalysis; D002470:Cell Survival / Q000187:drug effects; D057930:Click Chemistry; D003300:Copper; D061565:Cycloaddition Reaction; D005452:Fluoresceins / Q000737:chemistry / Q000633:toxicity; D057809:HEK293 Cells; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000737:chemistry* / Q000378:metabolism; D051379:Mice; D008856:Microscopy, Fluorescence; D041681:NIH 3T3 Cells; D011419:Propidium / Q000737:chemistry / Q000633:toxicity
chemical_list: D000480:Alkynes; D051017:Apoptosis Regulatory Proteins; D001386:Azides; D001672:Biocompatible Materials; D005452:Fluoresceins; D008562:Membrane Glycoproteins; D011419:Propidium; D003300:Copper; D006861:Hydrogen Peroxide; C018506:diacetylfluorescein
keywords: biocompatibility; click chemistry; copper-catalyzed azide-alkyne cycloaddition; glyco-labeling efficiency; glycoconjugates; strain-promoted azide-alkyne cycloaddition