Oncotically Driven Control over Glycocalyx Dimension for Cell Surface Engineering and Protein Binding in the Longitudinal Direction

Scientific Reports
Erika M J SirenJayachandran N Kizhakkedathu

Abstract

Here we present a simple technique for re-directing reactions on the cell surface to the outermost region of the glycocalyx. Macromolecular crowding with inert polymers was utilized to reversibly alter the accessibility of glycocalyx proteoglycans toward cell-surface reactive probes allowing for reactivity control in the longitudinal direction ('z'-direction) on the glycocalyx. Studies in HUVECs demonstrated an oncotically driven collapse of the glycocalyx brush structure in the presence of crowders as the mechanism responsible for re-directing reactivity. This phenomenon is consistent across a variety of macromolecular agents including polymers, protein markers and antibodies which all displayed enhanced binding to the outermost surface of multiple cell types. We then demonstrated the biological significance of the technique by increasing the camouflage of red blood cell surface antigens via a crowding-enhanced attachment of voluminous polymers to the exterior of the glycocalyx. The accessibility to Rhesus D (RhD) and CD47 proteins on the cell surface was significantly decreased in crowding-assisted polymer grafting in comparison to non-crowded conditions. This strategy is expected to generate new tools for controlled glycocal...Continue Reading

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Citations

Jun 10, 2020·Proceedings of the National Academy of Sciences of the United States of America·Sungmin SonDaniel A Fletcher
May 13, 2021·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Erika M J SirenJayachandran N Kizhakkedathu
Jun 3, 2021·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Dina BaierUrsula Windberger

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Methods Mentioned

BETA
confocal microscopy
flow cytometry

Software Mentioned

Graphpad Prism

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