Development of a monoclonal antibody against the canine CCR2 as a new targeted anticancer therapy in the scope of comparative oncology

Abstract

Event Abstract Back to Event Development of a monoclonal antibody against the canine CCR2 as a new targeted anticancer therapy in the scope of comparative oncology Teresa P. Raposo1*, David J. Argyle2, Breno C. Beirao2, Justina Prada1, Isabel Pires1 and Felisbina L. Queiroga1 1 University of Trás-os-Montes and Alto Douro, Department of Veterinary Sciences, Portugal 2 University of Edinburgh, The Royal (Dick) School of Veterinary Studies and The Roslin Institute, United Kingdom Introduction: In this project, it is proposed that the clinical application of a monoclonal antibody against the receptor of the Monocyte Chemotactic Protein-1 (MAb anti-CCR2), would hinder cancer progression, inhibiting macrophages recruitment. Canine mammary tumour (CMT) will be used as a model of human breast cancer, in face of molecular, histologic and clinical similarities of the disease in both species. Herein, the initial stages of the production of a MAb anti-CCR2 are described, aiming the future clinical application. Material and Methods: RNA was extracted from canine bone marrow, CCR2 cDNA was synthesized by RT-PCR, cloned into a plasmid vector, and then confirmed through DNA sequencing. TransformedBL21 DE3 E.coli were used as a system of bacterial expression. The bacterial lysates were purified through an affinity chromatography column, to isolate the CCR2 protein which was subsequently verified through mass spectrometry. Results: The purity of the isolated protein was confirmed by mass spectrometry analysis and it is now suitable for the mice immunisation. Throughout the purification process, the inclusion of urea in the lysis, wash and elution buffers was crucial to withdraw the most of the protein from the bacterial lysates. Conclusion: Hitherto, the antigen production stage was concluded, obtaining a pure form of CCR2, ready for the immunisation of mice.Hence, it is now possible to progress to the following phases in the development of a Mab anti-CCR2 using hybridoma technology. Afterwards, an experimental therapy delivering anti-CCR2 monoclonal antibody to the tumour site will be applied to dogs with advanced CMT. Acknowledgements Ministry of Science and Education, Portugal: QREN - POPH - FCT Project: SFRH / BD / 79158 / 2011 Keywords: tumour-associated macrophages, TAMs, canine cancer model, CCR2 antibody, MAb technology Conference: 15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013. Presentation Type: Abstract Topic: Translational immunology and immune intervention Citation: Raposo TP, Argyle DJ, Beirao BC, Prada J, Pires I and Queiroga FL (2013). Development of a monoclonal antibody against the canine CCR2 as a new targeted anticancer therapy in the scope of comparative oncology. Front. Immunol. Conference Abstract: 15th International Congress of Immunology (ICI). doi: 10.3389/conf.fimmu.2013.02.00269 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 12 Mar 2013; Published Online: 22 Aug 2013. * Correspondence: Miss. Teresa P Raposo, University of Trás-os-Montes and Alto Douro, Department of Veterinary Sciences, Vila Real, Portugal, rapteresa@gmail.com Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Teresa P Raposo David J Argyle Breno C Beirao Justina Prada Isabel Pires Felisbina L Queiroga Google Teresa P Raposo David J Argyle Breno C Beirao Justina Prada Isabel Pires Felisbina L Queiroga Google Scholar Teresa P Raposo David J Argyle Breno C Beirao Justina Prada Isabel Pires Felisbina L Queiroga PubMed Teresa P Raposo David J Argyle Breno C Beirao Justina Prada Isabel Pires Felisbina L Queiroga Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.