Characterization of H type 1 and type 1 N-acetyllactosamine glycan epitopes on ovarian cancer specifically recognized by the anti-glycan monoclonal antibody mAb-A4.

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Title: Characterization of H type 1 and type 1 N-acetyllactosamine glycan epitopes on ovarian cancer specifically recognized by the anti-glycan monoclonal antibody mAb-A4.
Author(s): Choo, M
Tan, HL
Ding, V
Castangia, R
Belgacem, O
Liau, B
Hartley-Tassell, L
Haslam, SM
Dell, A
Choo, A
Item Type: Journal Article
Abstract: Cancer-specific glycans of ovarian cancer are promising epitopes for targeting with monoclonal antibodies (mAb). Despite their potential, structural characterization of these glycan epitopes remains a significant challenge in mAb preclinical development. Our group generated the monoclonal antibody mAb-A4 against human embryonic stem cells (hESC), which also bound specifically to N-glycans present on 11 out of 19 ovarian cancer (OC) and 8 out of 14 breast cancer cell lines tested. Normal cell lines and tissue were unstained by mAb-A4. To characterize the N-linked glycan epitopes on OC cell lines targeted by mAb-A4, we used glycosidases, glycan microarray, siRNA and advanced high-sensitivity matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). The mAb-A4 epitopes were found to be Fuc α1-2Galβ1-3GlcNAcβ (H Type 1) and Galβ1-3GlcNAcβ (Type 1 LacNAc). These structures were found to be present on multiple proteins from hESC and OC. Importantly, endo-β-galactosidase coupled with MALDI-MS allowed these two epitopes, for the first time, to be directly identified on the polylactosamines of N-glycans of SKOV3, IGROV1, OV90 and OVCA433. Furthermore, siRNA knockdown of B3GALT5 expression in SKOV3 demonstrated that mAb-A4 binding was dependent on B3GALT5, providing orthogonal evidence of the epitopes' structures. The recognition of oncofetal H Type 1 and Type 1 LacNAc on OC by mAb-A4 is a novel and promising way to target OC and supports the theory that cancer can acquire stem-like phenotypes. We propose that the orthogonal framework used in this work could be the basis for advancing anti-glycan mAb characterization.
Publication Date: 6-Feb-2017
Date of Acceptance: 2-Feb-2017
URI: http://hdl.handle.net/10044/1/48284
DOI: https://dx.doi.org/10.1074/jbc.M116.768887
ISSN: 1083-351X
Publisher: American Society for Biochemistry and Molecular Biology
Start Page: 6163
End Page: 6176
Journal / Book Title: Journal of Biological Chemistry
Volume: 292
Issue: 15
Copyright Statement: © 2017 The American Society for Biochemistry and Molecular Biology, Inc. Final version free via Creative Commons CC-BY license (https://creativecommons.org/licenses/by/4.0/).
Sponsor/Funder: Biotechnology and Biological Sciences Research Council (BBSRC)
Funder's Grant Number: BB/K016164/1
Keywords: Science & Technology
Life Sciences & Biomedicine
Biochemistry & Molecular Biology
EMBRYONIC STEM-CELLS
LEWIS Y ANTIGEN
EPITHELIAL OVARIAN
ESCHERICHIA-FREUNDII
BIOSYNTHETIC-PATHWAY
CYTOTOXIC ANTIBODY
ANTITUMOR-ACTIVITY
MASS-SPECTROMETRY
GLYCOMIC ANALYSIS
DNA METHYLATION
cancer biology
cancer therapy
embryonic stem cell
glycobiology
glycoconjugate
glycomics
mass spectrometry (MS)
monoclonal antibody
ovarian cancer
small interfering RNA (siRNA)
Amino Sugars
Antibodies, Monoclonal, Murine-Derived
Antibodies, Neoplasm
Antigens, Neoplasm
Breast Neoplasms
Cell Line, Tumor
Epitopes
Female
Humans
Neoplastic Stem Cells
Ovarian Neoplasms
Biochemistry & Molecular Biology
06 Biological Sciences
11 Medical And Health Sciences
03 Chemical Sciences
Publication Status: Published
Conference Place: United States
Appears in Collections:Faculty of Natural Sciences



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