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Novel Carbohydrate Specificity of Monoclonal Antibody 91.9H Prepared against Human Colonic Sulfomucin: Recognition of Sulfo-Lewis a Structure

https://doi.org/10.1006/bbrc.1998.9726Get rights and content

Abstract

Monoclonal antibody (mAb) 91.9H was previously prepared against partially purified human colonic sulfomucins. The epitope was detected in normal colonic mucosa and primary and metastatic colorectal carcinoma in decreasing order of magnitude. In the present study, this antibody was shown to recognize sulfo-Leastructure, HSO3-3Galβ1-3(Fucα1-4)GlcNAc. Interactions between mAb 91.9H and synthetic oligosaccharides conjugated with biotinylated polyacrylamide carrier were examined by a biosensor based on surface plasmon resonance and by enzyme-linked immunosorbent assays. This mAb bound to sulfo-Leabut not to sulfo-LeX, Lea, LeX, sialyl-Lea, or sialyl-LeX. Sulfo-Leaoligosaccharides decreased its binding affinity with mAb 91.9H after periodate oxidation of its fucose residue. Immunohistochemical study showed a strong binding of mAb 91.9H to goblet cells in human colonic epithelia of Lewis-positive individuals but a trace binding in Lewis-negative individuals, confirming the specificity of this antibody toward structures containing a fucosylated type 1 backbone.

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    Abbreviations used: AAL,Aleuria aurantialectin; ABTS, 2, 2′-azinobis(3-ethylbenzthiazoline-sulfonic acid); BSA, bovine serum albumin; ELISA, enzyme-linked immunosorbent assay; Lea, Galβ1-3(Fucα1-4)GlcNAc; LeX, Galβ1-4(Fucα1-3)GlcNAc; mAb, monoclonal antibody; r. t., room temperature; SPR, surface plasmon resonance; sulfo-Lea, HSO3-3Galβ1-3(Fucα1-4)GlcNAc; sulfo-LeX, HSO3-3Galβ1-4(Fucα1-3)GlcNAc; TBS, Tris–HCl-buffered saline, pH 7.3; TBST, TBS containing 0.05% Tween 20

    1

    To whom correspondence should be addressed at Laboratory of Cancer Biology and Molecular Immunology, Graduate School of Pharmaceutical Sciences, University of Tokyo, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan. Fax: +81-3-3815-9344. E-mail:[email protected].

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