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Lectin inhibits antigen–antibody reaction in a glycoform‐specific manner: Application for detecting α2,6sialylated‐carcinoembryonic antigen
Author(s) -
Ito Hiromi,
Hoshi Kyoka,
Osuka Fumihiko,
Gotoh Mitsukazu,
Saito Takuro,
Hojo Hiroshi,
Suzuki Rei,
Ohira Hiromasa,
Honda Takashi,
Hashimoto Yasuhiro
Publication year - 2016
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.201600117
Subject(s) - carcinoembryonic antigen , lectin , agglutinin , antigen , peanut agglutinin , antibody , chemistry , wheat germ agglutinin , microbiology and biotechnology , glycan , colorectal cancer , adenocarcinoma , metastasis , glycoprotein , biology , biochemistry , cancer , immunology , genetics
Carcinoembryonic antigen (CEA) is a glycoprotein marker, which is widely used for diagnosing various cancers, especially colon adenocarcinoma. In addition, CEA mediates homotypic adhesion of colon adenocarcinoma cells, which appears to favor hematogenous metastasis. CEA carries α2,6sialyl residues on its N ‐glycans whereas a normal counterpart, normal fecal antigen‐2, does α2,3sialyl residues, suggesting that cancer‐specific α2,6sialylation on CEA may play a role for cell invasion and metastasis. A simple and rapid estimation of α2,6sialyled CEA in detergent extracts from formalin‐fixed colon adenocarcinoma by “lectin inhibition” is reported. In the lectin inhibition method, Sambucus sieboldiana Agglutinin (SSA) lectin, an α2,6sialic acid binder, was used as a glycoform‐specific inhibitor for antigen–antibody reaction in ELISA. Detergent extracts from colon adenocarcinoma showed a fair amount of ELISA signal in the absence of SSA whereas the signal was markedly reduced (45≈74%) in the presence of SSA, suggesting that the extracts contains α2,6sialyled CEA. The presence of α2,6sialyled CEA in the extracts was confirmed by lectin microarray, in which SSA, Sambucus nigra agglutinin, and Trichosanthes japonica agglutinin I lectins were used as α2,6sialyl binders. Thus lectin inhibition is a simple and rapid method for detecting α2,6sialyled CEA even in crude detergent extracts from formalin‐fixed adenocarcinoma tissue.