Rabbit derived VL single-domains as promising scaffolds to generate antibody-drug conjugates
Ana S. André University of Lisbon / Associate Laboratory for Animal and Veterinary Sciences
Frederico Aires-da-Silva University of Lisbon / Associate Laboratory for Animal and Veterinary Sciences
Luís Tavares University of Lisbon / Associate Laboratory for Animal and Veterinary Sciences
Solange Gil University of Lisbon / Associate Laboratory for Animal and Veterinary Sciences
Pedro M. P. Gois Universidade de Lisboa
Joao Goncalves Universidade de Lisboa
Miguel Castanho Universidade de Lisboa
João D. G. Correia Universidade de Lisboa
Lurdes Gano Universidade de Lisboa
University of Veterinary Medicine Vienna
Berta São Braz University of Lisbon / Associate Laboratory for Animal and Veterinary Sciences
Belmira Carrapiço University of Lisbon / Associate Laboratory for Animal and Veterinary Sciences
Gonçalo Vicente Universidade de Lisboa
Soraia Oliveira Technophage SA
Vera Neves Universidade de Lisboa
Marco Cavaco Universidade de Lisboa
João P M António Universidade de Lisboa
Joana Inês Carvalho University of Lisbon
Pedro Bule University of Lisbon / Associate Laboratory for Animal and Veterinary Sciences
Sara Nogueira University of Lisbon / Associate Laboratory for Animal and Veterinary Sciences
Sandra Aguiar University of Lisbon / Associate Laboratory for Animal and Veterinary Sciences
Joana N. R. Dias University of Lisbon / Associate Laboratory for Animal and Veterinary Sciences
Nature Portfolio
Antibody-drug conjugates (ADCs) are among the fastest-growing classes of therapeutics in oncology. Although ADCs are in the spotlight, they still present significant engineering challenges. Therefore, there is an urgent need to develop more stable and effective ADCs. Most rabbit light chains have an extra disulfide bridge, that links the variable and constant domains, between Cys80 and Cys171, which is not found in the human or mouse. Thus, to develop a new generation of ADCs, we explored the potential of rabbit-derived VL-single-domain antibody scaffolds (sdAbs) to selectively conjugate a payload to Cys80. Hence, a rabbit sdAb library directed towards canine non-Hodgkin lymphoma (cNHL) was subjected to in vitro and in vivo phage display. This allowed the identification of several highly specific VL-sdAbs, including C5, which specifically target cNHL cells in vitro and present promising in vivo tumor uptake. C5 was selected for SN-38 site-selective payload conjugation through its exposed free Cys80 to generate a stable and homogenous C5-DAB-SN-38. C5-DAB-SN-38 exhibited potent cytotoxicity activity against cNHL cells while inhibiting DNA-TopoI activity. Overall, our strategy validates a platform to develop a novel class of ADCs that combines the benefits of rabbit VL-sdAb scaffolds and the canine lymphoma model as a powerful framework for clinically translation of novel therapeutics for cancer.
Englisch
2023
Dieses Werk bzw. dieser Inhalt steht unter einer
CC BY 4.0 - Creative Commons Namensnennung 4.0 International Lizenz.
CC BY 4.0 International
http://creativecommons.org/licenses/by/4.0/
Therapeutic Activity; Next-Generation; Strategies; Selection; Peptide