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    An assay for quantitative analysis of polysialic acid expression in cancer cells

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    Publication date
    2021-05-01
    Author
    Guo, Xiaoxiao
    Elkashef, Sara M.
    Patel, Anjana
    Ribeiro Morais, Goreti
    Shnyder, Steven D.
    Loadman, Paul M.
    Patterson, Laurence H.
    Falconer, Robert A.
    Keyword
    Polysialic acid
    Sialic acid
    Quantitative analysis
    Cell-based method
    Inhibitor screening
    Rights
    © 2021 Elsevier. Reproduced in accordance with the publisher's self-archiving policy. This manuscript version is made available under the CC-BY-NC-ND 4.0 license (https://creativecommons.org/licenses/by-nc-nd/4.0/)
    Peer-Reviewed
    Yes
    Open Access status
    Green
    
    Metadata
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    Abstract
    Polysialic acid (polySia) is a linear polysaccharide comprised of N-acetylneuraminic acid residues and its over-expression in cancer cells has been correlated with poor clinical prognosis. An assay has been developed for quantitative analysis of cellular polySia expression. This was achieved by extracting and purifying released polySia from glycoproteins by mild acid hydrolysis and optimised organic extraction. The polySia was further hydrolysed into Sia monomers, followed by fluorescent labelling and quantitative analysis. The assay was qualified utilising endoneuraminidase-NF to remove polySia from the surface of C6-ST8SiaII cancer cells (EC50 = 2.13 ng/ml). The result was comparable to that obtained in a polySia-specific cellular ELISA assay. Furthermore, the assay proved suitable for evaluation of changes in polySia expression following treatment with a small molecule inhibitor of polysialylation. Given the importance of polySia in multiple disease states, notably cancer, this is a potentially vital tool with applications in the fields of drug discovery and glycobiology.
    URI
    http://hdl.handle.net/10454/18344
    Version
    Accepted manuscript
    Citation
    Guo X, Elkashef SM, Patel A et al (2021) An assay for quantitative analysis of polysialic acid expression in cancer cells. Carbohydrate Polymers. 259: 117741.
    Link to publisher’s version
    https://doi.org/10.1016/j.carbpol.2021.117741
    Type
    Article
    Collections
    Life Sciences Publications

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