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Sci Rep. 2018 Aug 15;8(1):12196. doi: 10.1038/s41598-018-30421-w.

Novel monoclonal antibody L2A5 specifically targeting sialyl-Tn and short glycans terminated by alpha-2-6 sialic acids.

Author information

1
UCIBIO-REQUIMTE, Department of Life Sciences, Faculty of Science and Technology, NOVA University of Lisbon, Lisbon, 2829, Portugal.
2
iBET, Instituto de Biologia Experimental e Tecnológica, Oeiras, 2780, Portugal.
3
Experimental Pathology and Therapeutics Group, IPO-Porto Research Center, Portuguese Institute of Oncology of Porto, Porto, 4200, Portugal.
4
Glycosciences Laboratory - Department of Medicine, Imperial College London, London, W12 0NN, United Kingdom.
5
Glycobiology in Cancer, Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP), Porto, 4200, Portugal.
6
Institute for Research and Innovation in Health (I3S), University of Porto, 4200, Porto, Portugal.
7
CINTESIS - Center for Health Technology and Services Research, University of Porto, Porto, 4200, Portugal.
8
Molecular Oncology and Viral Pathology Group, IPO-Porto Research Center, Portuguese Oncology Institute of Porto, Porto, 4200, Portugal.
9
Joaquim Chaves Saúde, Anatomical Pathology Laboratory, Lisboa, 1170, Portugal.
10
UCIBIO-REQUIMTE, Department of Chemistry, Faculty of Science and Technology, NOVA University of Lisbon, Lisbon, 2829, Portugal.
11
Key Laboratory of Marine Drugs of Ministry of Education, and Shandong Provincial Key Laboratory of Glycoscience and Glycoengineering, School of Medicine and Pharmacy, Ocean University of China, Qingdao, 266003, China.
12
Institute of Biomedical Sciences Abel Salazar, University of Porto, Porto, 4050, Portugal.
13
Department of Surgical Oncology, Portuguese Institute of Oncology, Porto, 4200, Portugal.
14
International Iberian Nanotechnology Laboratory (INL), Braga, 4715, Portugal.
15
Bayer Portugal, Carnaxide, 2790, Portugal.
16
UCIBIO-REQUIMTE, Department of Life Sciences, Faculty of Science and Technology, NOVA University of Lisbon, Lisbon, 2829, Portugal. cnovo@ihmt.unl.pt.
17
UEIPM, Institute of Hygiene and Tropical Medicine, NOVA University of Lisbon, Lisbon, 1349, Portugal. cnovo@ihmt.unl.pt.
18
UCIBIO-REQUIMTE, Department of Life Sciences, Faculty of Science and Technology, NOVA University of Lisbon, Lisbon, 2829, Portugal. p.videira@fct.unl.pt.
19
CDG & Allies - Professionals and Patient Associations International Network (CDG & Allies - PPAIN), Caparica, 2829, Portugal. p.videira@fct.unl.pt.

Abstract

Incomplete O-glycosylation is a feature associated with malignancy resulting in the expression of truncated glycans such as the sialyl-Tn (STn) antigen. Despite all the progress in the development of potential anti-cancer antibodies, their application is frequently hindered by low specificities and cross-reactivity. In this study, a novel anti-STn monoclonal antibody named L2A5 was developed by hybridoma technology. Flow cytometry analysis showed that L2A5 specifically binds to sialylated structures on the cell surface of STn-expressing breast and bladder cancer cell lines. Moreover, immunoblotting assays demonstrated reactivity to tumour-associated O-glycosylated proteins, such as MUC1. Tumour recognition was further observed using immunohistochemistry assays, which demonstrated a high sensitivity and specificity of L2A5 mAb towards cancer tissue, using bladder and colorectal cancer tissues. L2A5 staining was exclusively tumoural, with a remarkable reactivity in invasive and metastasis sites, not detectable by other anti-STn mAbs. Additionally, it stained 20% of cases of triple-negative breast cancers, suggesting application in diseases with unmet clinical needs. Finally, the fine specificity was assessed using glycan microarrays, demonstrating a highly specific binding of L2A5 to core STn antigens and additional ability to bind 2-6-linked sialyl core-1 probes. In conclusion, this study describes a novel anti-STn antibody with a unique binding specificity that can be applied for cancer diagnostic and future development of new antibody-based therapeutic applications.

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