Quantitative Screening of Cell‐Surface Gangliosides by Nondestructive Extraction and Hydrophobic Collection

Abstract

As creening strategy involving designed extractors and collectors was used for the nondestructive quantitation of gangliosides on cell surfaces.T he extractors were constructed by functionalizing maleimide silica bubbles with aDNA probe, which contains an endonuclease cleavage site and ab oronic acid end to extract cell-surface sialic acid-containing compounds through simple centrifugation. After the extractors containing the extracted compounds were incubated with endonuclease,t he released oligonucleotide-gangliosides were selectively collected by silanized collector bubbles through hydrophobic interactions.The in vitro fluorescent signals from the collectors were used for the quantitation of cell-surface gangliosides.Bycombining with sialidase cleavage,aprotocol for the identification of ganglioside subtypes was developed. The successful monitoring of the regeneration of cell-surface gangliosides demonstrates the potential of this strategy in probing related biological processes.

In Situ Cellular Glycan Analysis

“…By modifying PBA onto nanostructures such as gold nanoflowers 22 or nanoclusters, 23 cell surface Sia can be sensitively detected by SERS or fluorescence. In addition, Sia-contained glycolipids, i.e., gangliosides, 24 can also be quantitated with a PBA functionalized silica bubble. Metabolic labeling is based on incorporation of a bioorthogonal chemical reporter into a target biomolecule using the biosynthetic machinery of cells, 25 which does not produce significant structural perturbation and can be tolerated by promiscuous enzymes involved in the installation process.…”