BibTex format
@article{Choo:2017:10.1074/jbc.M116.768887,
author = {Choo, M and Tan, HL and Ding, V and Castangia, R and Belgacem, O and Liau, B and Hartley-Tassell, L and Haslam, SM and Dell, A and Choo, A},
doi = {10.1074/jbc.M116.768887},
journal = {Journal of Biological Chemistry},
pages = {6163--6176},
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.},
url = {http://dx.doi.org/10.1074/jbc.M116.768887},
volume = {292},
year = {2017}
}
RIS format (EndNote, RefMan)
TY - JOUR
AB - 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.
AU - Choo,M
AU - Tan,HL
AU - Ding,V
AU - Castangia,R
AU - Belgacem,O
AU - Liau,B
AU - Hartley-Tassell,L
AU - Haslam,SM
AU - Dell,A
AU - Choo,A
DO - 10.1074/jbc.M116.768887
EP - 6176
PY - 2017///
SN - 1083-351X
SP - 6163
TI - 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.
T2 - Journal of Biological Chemistry
UR - http://dx.doi.org/10.1074/jbc.M116.768887
UR - http://www.ncbi.nlm.nih.gov/pubmed/28167527
VL - 292
ER -