Structure-guided phage display discovery of antibodies for (S)Tn-glycans in protein context

Hurtado-Guerrero, Ramón ; Spyridon, Gatos ; Ginés-Alcober, Irene ; Macías-León, Javier ; González-Ramírez, Andrés Manuel ; Kasapoglu, Ilknur ; Veloz, Billy (Universidad de Zaragoza) ; Compañón, Ismael ; Ghirardello, Mattia ; Merino, Pedro (Universidad de Zaragoza) ; Corzana, Francisco ; Blixt, Ola

Resumen: Developing high-affinity monoclonal antibodies (mAbs) against tumor-associated carbohydrate antigens such as Tn and STn on carrier proteins remains a major challenge in cancer therapy. These antigens, expressed as glycan–peptide epitopes (combotopes), require precise recognition for high specificity. Through structural studies, we found that VH domains of certain antibodies primarily recognize glycans, whereas VL domains bind peptide sequences. Using these insights, we developed a VH-focused and VL-diverse phage display library to discover mAbs with combotope-binding characteristics. Notably, structural analysis enabled us to convert Tn-specific mAbs into STn-specific mAbs through modification of VH complementarity-determining region 3, demonstrating the versatility of this approach. Our hypothesis was validated with glycoprotein targets MUC1 and CD43, yielding antibodies with high specificity and affinity. Furthermore, internalization studies using the parental antibody scaffold show efficient uptake by tumor cells, supporting its use in antibody–drug conjugates. This platform addresses the challenge of generating glycoform-specific antibodies for cancer therapy.

Idioma: Inglés

DOI: 10.1038/s41589-025-01971-8

Año: 2025

Publicado en: Nature Chemical Biology (2025), [18 pp.]
ISSN: 1552-4450

Financiación: info:eu-repo/grantAgreement/ES/AEI/PID2019-105451GB-I00
Financiación: info:eu-repo/grantAgreement/ES/DGA/E34-23R
Financiación: info:eu-repo/grantAgreement/ES/DGA-FEDER/E34-R17
Financiación: info:eu-repo/grantAgreement/ES/DGA/LMP58-18
Financiación: info:eu-repo/grantAgreement/EC/FP7/316929/EU/Joined efforts to fight gastric cancer/GastricGlycoExplorer
Financiación: info:eu-repo/grantAgreement/EC/H2020/814029/EU/Synthetic biology of carbohydrate-binding proteins: engineering protein-carbohydrate interactions for diagnostics and cell targeting
Financiación: info:eu-repo/grantAgreement/ES/MICINN AEI/PID2022-136362NB-I00
Financiación: info:eu-repo/grantAgreement/ES/MICINN/PID2021-127622OB-I00
Financiación: info:eu-repo/grantAgreement/ES/MICINN/PID2022-137973NB-I00

Tipo y forma: Artículo (Versión definitiva)

Área (Departamento): Área Química Orgánica (Dpto. Química Orgánica)
Área (Departamento): Área Bioquímica y Biolog.Mole. (Dpto. Bioq.Biolog.Mol. Celular)