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Synthesis of bodipy-labeled bacterial polysaccharides and their interaction with human dendritic cells

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Abstract

In this report, we describe the fluorescent labeling of bacterial polysaccharides (Escherichia coli O86:B7, Escherichia coli O19ab, Pseudomonas aeruginosa O10a10b, and Shigella flexneri 2b) at the “natural” amino group of their phosphoethanolamine moiety. Two protocols for labeling are compared: 1) on a scale of a few mg of the polysaccharide, with a dialysis procedure for purification from excessive reagents; and 2) on a scale of 0.1 mg of the polysaccharide, with a simple precipitation procedure instead of dialysis. The microscale version is sufficient for comfortable cytofluorometric analysis. The resulting probes were found to specifically bind to human dendritic cells in a dose-dependent manner. The used limited set of polysaccharides did not allow us even to get close to understanding which dendritic cell-associated lectins and which cognate polysaccharide epitopes are involved in recognition, but the proposed microscale protocol allows to generate a library of fluorescent probes for further mapping of the polysaccharide specificity of the dendritic cells.

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Abbreviations

BSA:

Bovine serum albumin

DC:

Dendritic cell

DMSO:

Dimethylsulfoxide

LPS:

Lipopolysaccharide

PBS:

Phosphate buffer saline

PBA:

PBS containing 0.2% BSA

PS:

Polysaccharide

Su:

Succinimide

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Acknowledgments

This work was generously supported by grant 20-63-47110 of theRussian Science Foundation, and in part by grant 16-04-01084 of the Russian Foundation for Basic Research.

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Authors and Affiliations

  1. Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry RAS, 16/10 Miklukho-Maklaya str, Moscow, 117997, Russia

    Alexander B. Tuzikov, Eugenia M. Rapoport, Sergey V. Khaidukov, Elena A. Nokel & Nicolai V. Bovin

  2. Zelinsky Institute of Organic Chemistry RAS, 47 Leninsky prosp, Moscow, 119991, Russia

    Yuriy A. Knirel

  3. School of Engineering, Computer and Mathematical Sciences, Auckland University of Technology, Auckland, New Zealand

    Nicolai V. Bovin

Authors
  1. Alexander B. Tuzikov
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  2. Eugenia M. Rapoport
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  3. Sergey V. Khaidukov
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  4. Elena A. Nokel
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  5. Yuriy A. Knirel
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  6. Nicolai V. Bovin
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Correspondence to Nicolai V. Bovin.

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Tuzikov, A.B., Rapoport, E.M., Khaidukov, S.V. et al. Synthesis of bodipy-labeled bacterial polysaccharides and their interaction with human dendritic cells. Glycoconj J 38, 369–374 (2021). https://doi.org/10.1007/s10719-021-09993-9

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  • DOI: https://doi.org/10.1007/s10719-021-09993-9

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