lectin inhibits Antigen Antibody Reaction in a glycoform specific manner application for detecting α2 6sialylated carcinoembryonic Antigen

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 faecal Antigen-2, does α2,3sialyl residues, suggesting that cancer specific  α2,6sialylation on CEA may play a role for cell invasion and metastasis. Here we report a simple and rapid estimation of α2,6sialyled-CEA in detergent extracts from formalin fixed colon adenocarcinoma by “lectin inhibition”. 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.

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Lectin inhibits Antigen–Antibody Reaction in a glycoform‐specific manner: Application for detecting α2,6sialylated‐carcinoembryonic Antigen

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 faecal Antigen-2, does α2,3sialyl residues, suggesting that cancer specific  α2,6sialylation on CEA may play a role for cell invasion and metastasis. Here we report a simple and rapid estimation of α2,6sialyled-CEA in detergent extracts from formalin fixed colon adenocarcinoma by “lectin inhibition”. 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.

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lectin dependent inhibition of Antigen Antibody Reaction application for measuring α2 6 sialylated glycoforms of transferrin

We developed a high-throughput Enzyme-linked immunosorbent assay (ELISA) for measuring α2,6-sialylated transferrin (Tf), based on inhibition of anti-Tf Antibody binding to α2,6-sialylated Tf by a lectin, Sambucus sieboldiana Agglutinin (SSA). The 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. This finding was applied to an automated latex-agglutination immunoassay, using SSA lectin as an inhibitor (SSA-ALI). The concentration of α2,6-sialylated Tf measured by SSA-ALI in human cerebrospinal fluid was correlated with that of ELISA (r2 = 0.8554), previously developed for measuring α2,6-sialylated Tf.

lectin inhibits Antigen Antibody Reaction in a glycoform specific manner application for detecting α2 6sialylated carcinoembryonic Antigen

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 faecal Antigen-2, does α2,3sialyl residues, suggesting that cancer specific  α2,6sialylation on CEA may play a role for cell invasion and metastasis. Here we report a simple and rapid estimation of α2,6sialyled-CEA in detergent extracts from formalin fixed colon adenocarcinoma by “lectin inhibition”. 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.

This article is protected by copyright. All rights reserved

Lectin inhibits Antigen–Antibody Reaction in a glycoform‐specific manner: Application for detecting α2,6sialylated‐carcinoembryonic Antigen

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 faecal Antigen-2, does α2,3sialyl residues, suggesting that cancer specific  α2,6sialylation on CEA may play a role for cell invasion and metastasis. Here we report a simple and rapid estimation of α2,6sialyled-CEA in detergent extracts from formalin fixed colon adenocarcinoma by “lectin inhibition”. 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.

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microfabricated renewable beads trapping releasing flow cell for rapid Antigen Antibody Reaction in chemiluminescent immunoassay

A renewable flow cell integrating a microstructured pillar-array filter and a pneumatic microvalve was microfabricated to trap and release beads. A bead-based immunoassay using this device was also developed. This microfabricated device consists of a microfluidic channel connecting to a beads chamber in which the pillar-array filter is built. Underneath the filter, there is a pneumatic microvalve built across the chamber. Such a device can trap and release beads in the chamber by “closing” or “opening” the microvalve. On the basis of the pneumatic microvalve, the device can trap beads in the chamber before performing an assay and release the used beads after the assay. Therefore, this microfabricated device is suitable for “renewable surface analysis”. A model analyte, 3,5,6-trichloropyridinol (TCP), was chosen to demonstrate the analytical performance of the device. The entire fluidic assay process, including beads trapping, immuno binding, beads washing, beads releasing, and chemiluminesence signal coll…