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Updated: Jul 28, 2026
Chemically-blocked Antibody Microarray for Multiplexed High-throughput Profiling of Specific Protein Glycosylation in Complex Samples
Published on: May 4, 2012
Identification of the O-Glycan Epitope Targeted by the Anti-Human Carcinoma Monoclonal Antibody (mAb) NEO-201
Kwong Y Tsang1, Massimo Fantini1, Anjum Zaki1
1Precision Biologics, Inc., Bethesda, MD 20814, USA.
Abstract:
Truncated O-glycans expressed in cancer cells support tumor progression, and they may serve as potential targets to improve the monitoring and treatment of cancers. Previously, we reported that NEO-201 binds to several tumors expressing tumor-associated CEACAM5 and CEACAM6 variants but does not bind to those expressed in healthy tissues. This specific binding may be associated with the presence of truncated O-glycans attached on the protein sequence of these variants. To evaluate the glycosylation pattern targeted by NEO-201 we performed an O-glycan array consisting of 94 O-glycans. O-glycan profiles were elucidated from the human pancreatic cancer cell line CFPAC-1, human hematological neoplastic cells (HL-60, U937, K562) and human neutrophils. The O-glycan array analysis showed that NEO-201 interacts with core 1-4 O-glycans and that the binding to a specific core 1 O-glycan was the strongest. The O-glycan profiling of the NEO-201-reactive cells CFPAC-1, HL-60, U937 and human neutrophils showed that cells recognized by NEO-201 express mostly core 1 and/or extended core 1 O-glycans. In addition, NEO-201 mediates antibody-dependent cell-mediated cytotoxicity (ADCC) against tumor cells expressing core 1 or extended core 1 O-glycan profiles. These results demonstrated that NEO-201 binds to core 1 and extended core 1 O-glycans expressed in its target cells. Since GalNAc residue can be added onto threonine and serine to form O-glycans, it is very likely that NEO-201 recognizes these O-glycans attached to any protein with amino acid regions containing serine and threonine.
Insights
NEO-201 antibody targets truncated O-glycans, specifically core 1 and extended core 1 structures found on cancer cells. This binding facilitates antibody-dependent cell-mediated cytotoxicity, offering potential for cancer monitoring and treatment.
Area of Science:
- Glycoscience
- Cancer Biology
- Immunology
Background:
- Truncated O-glycans on cancer cells promote tumor progression and are potential therapeutic targets.
- NEO-201 antibody shows specific binding to tumor-associated CEACAM5 and CEACAM6 variants, potentially due to these O-glycans.
Purpose of the Study:
- To evaluate the specific O-glycan structures targeted by the NEO-201 antibody.
- To investigate the O-glycan profiles of cancer cells recognized by NEO-201.
- To assess the therapeutic potential of NEO-201 in mediating antibody-dependent cell-mediated cytotoxicity (ADCC).
Main Methods:
- O-glycan array analysis using 94 different O-glycans.
- O-glycan profiling of human pancreatic cancer cell line (CFPAC-1), hematological neoplastic cells (HL-60, U937, K562), and human neutrophils.
- Assessment of NEO-201's ability to mediate ADCC against tumor cells.
Main Results:
- NEO-201 demonstrated interaction with core 1-4 O-glycans, with strongest binding to a specific core 1 O-glycan.
- NEO-201-reactive cells (CFPAC-1, HL-60, U937, neutrophils) predominantly express core 1 and/or extended core 1 O-glycans.
- NEO-201 effectively mediated ADCC against tumor cells expressing core 1 or extended core 1 O-glycan profiles.
Conclusions:
- NEO-201 specifically binds to core 1 and extended core 1 O-glycans.
- These findings suggest NEO-201 recognizes N-acetylgalactosamine (GalNAc) residues forming O-glycans on serine and threonine residues.
- NEO-201's ability to induce ADCC against cancer cells expressing these O-glycans highlights its potential as a cancer therapeutic and diagnostic agent.
