An improved synthesis of dansylated α-galactosylceramide and its use as a fluorescent probe for the monitoring of glycolipid uptake by cells

A highly efficient synthesis of the biologically important fluorescent probe dansyl α-GalCer is presented. Key in our strategy is the incorporation of the fluorescent dansyl group at an early stage in the synthesis to facilitate in the monitoring and purification of intermediates via TLC and flash column chromatography, respectively, and the use of a high yielding α-selective glycosylation reaction between the phytosphingosine lipid and a galactosyl iodide donor. The ability of dansyl α-GalCer to activate iNKT cells and to serve as a fluorescent marker for the uptake of glycolipid by dendritic cells is also presented.

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► An efficient and novel synthesis of a fluorescent α-GalCer probe was developed.
► Early incorporation of the dansyl fluorosphore greatly facilitated the synthesis.
► The dansylated probe activated iNKT cells in a CD1d dependent manner.
► Cells that took up dansyl α-GalCer were detected by flow cytometry.