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

Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry
Hiromi ItoYasuhiro Hashimoto

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 coloni...Continue Reading

References

Mar 10, 2001·Journal of the Neurological Sciences·H Reiber, J B Peter
Dec 14, 2007·Nature·Benjamin F CravattJohn R Yates
Jun 10, 2010·Nature Reviews. Genetics·R David HawkinsBing Ren
Jun 26, 2010·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Atsushi MatsudaHisashi Narimatsu
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Apr 5, 2011·Neurobiology of Aging·Satoshi FutakawaYasuhiro Hashimoto

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Citations

Sep 3, 2021·Insect Science·Pengyu ChenGuy Smagghe

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Methods Mentioned

BETA
glycosylation
electrophoresis
nuclear magnetic resonance
enzyme-linked immunosorbent assay
ELISA
Assay

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