Increasing the Efficacy of Bioorthogonal Click Reactions for Bioconjugation: A Comparative Study

Besanceney-Webler, Christen; Jiang, Hao; Zheng, Tianqing; Feng, Lei; Amo, David Soriano del; Wang, Wei; Klivansky, Liana M.; Marlow, Florence; Liu, Yi; Wu, Peng

doi:10.1002/ange.201101817

Cited by 137 publications

(135 citation statements)

References 36 publications

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“…In other words, the Ac 4 ManNAz feeding was performed at 0 d in vitro (DIV), and the CuAAC reaction was performed at 2 DIV. For the CuAAC reaction, a recently developed ligand, 2-[4-{{bis[(1-tert-butyl-1H-1,2,3-triazol-4-yl)methyl]amino}methyl}-1H-1,2,3-triazol-1-yl]acetic acid (36)(37)(38), was used to minimize the cytotoxicity of Cu(I) ions and fasten the coupling reaction (39). After a 5-min reaction at 4°C, the intense red fluorescence was observed at the entire cellular surface, including neuronal cell bodies (somas) and neurites ( Fig.…”

Section: Resultsmentioning

confidence: 99%

Tissue-based metabolic labeling of polysialic acids in living primary hippocampal neurons

Kang

Joo

Choi

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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The posttranslational modification of neural cell-adhesion molecule (NCAM) with polysialic acid (PSA) and the spatiotemporal distribution of PSA-NCAM play an important role in the neuronal development. In this work, we developed a tissue-based strategy for metabolically incorporating an unnatural monosaccharide, peracetylated N-azidoacetyl-D-mannosamine, in the sialic acid biochemical pathway to present N-azidoacetyl sialic acid to PSA-NCAM. Although significant neurotoxicity was observed in the conventional metabolic labeling that used the dissociated neuron cells, neurotoxicity disappeared in this modified strategy, allowing for investigation of the temporal and spatial distributions of PSA in the primary hippocampal neurons. PSA-NCAM was synthesized and recycled continuously during neuronal development, and the two-color labeling showed that newly synthesized PSANCAMs were transported and inserted mainly to the growing neurites and not significantly to the cell body. This report suggests a reliable and cytocompatible method for in vitro analysis of glycans complementary to the conventional cell-based metabolic labeling for chemical glycobiology.

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Section: Resultsmentioning

confidence: 99%

Tissue-based metabolic labeling of polysialic acids in living primary hippocampal neurons

Kang

Joo

Choi

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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“…6 Another approach is to design ligands, which coordinate to Cu I and enhance its biocompatibility. Typical ligands include bathophenanthroline disulfonate disodium salt (BPS), 7 14,41 tris((1-benzyl-1H-1,2,3-triazol-4-yl)methyl)amine (TBTA), 8 tris-(hydroxypropyltriazolyl)-methylamine (THPTA), 9 bis[(tertbutyltriazoyl) methyl]-[(2-carboxymethyltriazoyl)methyl]amine (BTTAA), 10 and L-histidine. 11 Despite promising results, it is still challenging to design biocompatible and biodegradable ligands that show excellent catalytic activity in complex living environment with minimal Cu I cytotoxicity.…”

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confidence: 99%

“…Previously, catalyst activity was usually tested in a mixture of organic solvents and water utilized for labeling of biomolecules. 10,11,18 Unlike those studies, we tested the activity of the supramolecular PA-Cu II nanofibers under physiological conditions to obtain more realistic results. A typical Huisgen 1,3-dipolar cycloaddition reaction was designed where phenylacetylene and benzylazide were chosen as reactants, sodium ascorbate as reducing agent, and water as biological solvent (Table1).…”

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confidence: 99%

Biocompatible Supramolecular Catalytic One-Dimensional Nanofibers for Efficient Labeling of Live Cells

et al. 2015

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Understanding complex cellular functions requires study and tracking of biomolecules such as proteins, glycans, and lipids in their natural environment. Herein, we report the first supramolecular nanocatalyst for bioorthogonal click reaction to label live cells. This biocompatible and biodegradable nanocatalyst was formed by self-assembled peptide nanofibers complexed with copper ions. The supramolecular nanocatalyst enhanced azide−alkyne cycloaddition reaction rate under physiological conditions and was shown to be useful for efficient bioorthogonal labeling of live cells.

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“…We microinjected embryos with 5 pmol BCNSia and/or 50 pmol SiaNAl, the expected metabolic product of an N-pentynoyl mannosamine derivative that we have previously described. [28] At 48 hpf, the embryos were injected with 4 and then bathed in a copper click solution containing sodium ascorbate, BTTAA ligand, [29] and the azide probe CalFluor 647 for 30 min. [28b] We observed robust BCNSiadependent labeling of interior cells and SiaNAl-dependent labeling of enveloping layer cells ( Figure 6B).…”

mentioning

confidence: 99%

Systemic Fluorescence Imaging of Zebrafish Glycans with Bioorthogonal Chemistry

et al. 2015

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Vertebrate glycans constitute a large, important, and dynamic set of post-translational modifications that are notoriously difficult to manipulate and image. Although the chemical reporter strategy has been used in conjunction with bioorthogonal chemistry to image the external glycosylation state of live zebrafish and detect tumor-associated glycans in mice, the ability to image glycans systemically within a live organism has remained elusive. Here, we report a method that combines the metabolic incorporation of a cyclooctyne-functionalized sialic acid derivative with a ligation reaction of a fluorogenic tetrazine, allowing for the imaging of sialylated glycoconjugates within live zebrafish embryos.

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Increasing the Efficacy of Bioorthogonal Click Reactions for Bioconjugation: A Comparative Study

Cited by 137 publications

References 36 publications

Tissue-based metabolic labeling of polysialic acids in living primary hippocampal neurons

Tissue-based metabolic labeling of polysialic acids in living primary hippocampal neurons

Biocompatible Supramolecular Catalytic One-Dimensional Nanofibers for Efficient Labeling of Live Cells

Systemic Fluorescence Imaging of Zebrafish Glycans with Bioorthogonal Chemistry

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