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Molecules 2018, 23(6), 1314; https://doi.org/10.3390/molecules23061314

Lectin-Based Assay for Glycoform-Specific Detection of α2,6-sialylated Transferrin and Carcinoembryonic Antigen in Tissue and Body Fluid

1
Department of Biochemistry, Fukushima Medical University School of Medicine, Fukushima 960-1295, Japan
2
Devision of Human Life Science, Fukushima Medical University School of Nursing, Fukushima 960-1295, Japan
These authors were equally contributed to the present study.
*
Author to whom correspondence should be addressed.
Received: 22 April 2018 / Revised: 16 May 2018 / Accepted: 25 May 2018 / Published: 30 May 2018
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Abstract

Antibodies are useful for detecting glycoprotein antigens, but a conventional antibody recognizes only a protein epitope rather than a glycan. Thus, glycan isoform detection generally requires time- and labor-consuming processes such as lectin affinity column chromatography followed by sandwich ELISA. We recently found antigen-antibody reactions that were inhibited by lectin binding to glycans on the glycoprotein antigen, leading to a convenient glycoform-specific assay. Indeed, Sambucus sieboldiana agglutinin (SSA) lectin, a binder to sialylα2,6galactose residue, inhibited antibody binding to α2,6-sialylated transferrin (Tf) (SSA inhibition). SSA inhibition was not observed with other glycoforms, such as periodate-treated, sialidase-treated and sialidase/galactosidase-treated Tf, suggesting that the assay was glycoform-specific. SSA inhibition was also applicable for visualizing localization of α2,6-sialylated-Tf in a liver section. This is the first immunohistochemical demonstration of glycoform localization in a tissue section. SSA inhibition was utilized for establishing ELISA to quantify α2,6-sialylated carcinoembryonic antigen (CEA), a marker for various cancers. In addition, α2,6-sialylated-CEA was visualized in a colonic adenocarcinoma section by SSA inhibition. The method would further be applicable to a simple and rapid estimation of other α2,6-sialylated glycoproteins and have a potential aid to histopathological diagnosis. View Full-Text
Keywords: adenocarcinoma; antigen-antibody reaction; automated latex-agglutination immunoassay; carcinoembryonic antigen; cerebrospinal fluid; immunohistochemistry; enzyme-linked immunosorbent assay; Psathyrella velutina lectin; Sambucus sieboldiana agglutinin; transferrin; sialylα2,6galactose adenocarcinoma; antigen-antibody reaction; automated latex-agglutination immunoassay; carcinoembryonic antigen; cerebrospinal fluid; immunohistochemistry; enzyme-linked immunosorbent assay; Psathyrella velutina lectin; Sambucus sieboldiana agglutinin; transferrin; sialylα2,6galactose
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).
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Ito, H.; Hoshi, K.; Honda, T.; Hashimoto, Y. Lectin-Based Assay for Glycoform-Specific Detection of α2,6-sialylated Transferrin and Carcinoembryonic Antigen in Tissue and Body Fluid. Molecules 2018, 23, 1314.

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