title: Generation of a canine anti-EGFR (ErbB-1) antibody for passive immunotherapy in dog cancer patients.
abstract: Passive immunotherapy with monoclonal antibodies represents a cornerstone of human anticancer therapies, but has not been established in veterinary medicine yet. As the tumor-associated antigen EGFR (ErbB-1) is highly conserved between humans and dogs, and considering the effectiveness of the anti-EGFR antibody cetuximab in human clinical oncology, we present here a "caninized" version of this antibody, can225IgG, for comparative oncology studies. Variable region genes of 225, the murine precursor of cetuximab, were fused with canine constant heavy gamma and kappa chain genes, respectively, and transfected into Chinese hamster ovary (CHO) DUKX-B11 cells. Of note, 480 clones were screened and the best clones were selected according to productivity and highest specificity in EGFR-coated ELISA. Upon purification with Protein G, the recombinant cetuximab-like canine IgG was tested for integrity, correct assembly, and functionality. Specific binding to the surface of EGFR-overexpressing cells was assessed by flow cytometry and immunofluorescence; moreover, binding to canine mammary tissue was demonstrated by immunohistochemistry. In cell viability and proliferation assays, incubation with can225IgG led to significant tumor cell growth inhibition. Moreover, this antibody mediated significant tumor cell killing via phagocytosis in vitro. We thus present here, for the first time, the generation of a canine IgG antibody and its hypothetical structure. On the basis of its cetuximab-like binding site, on the one hand, and the expression of a 91% homologous EGFR molecule in canine cancer, on the other hand, this antibody may be a promising research compound to establish passive immunotherapy in dog patients with cancer.
authors.affiliation: Comparative Immunology and Oncology, Institute of Pathophysiology and Allergy Research, Medical University of Vienna.|Comparative Medicine, Messerli Research Institute, University of Veterinary Medicine Vienna, Medical University Vienna and University Vienna.
authors.forename: Josef
authors.initials: J
authors.lastname: Singer
authors.affiliation: Comparative Immunology and Oncology, Institute of Pathophysiology and Allergy Research, Medical University of Vienna.|Comparative Medicine, Messerli Research Institute, University of Veterinary Medicine Vienna, Medical University Vienna and University Vienna.|Department for Applied Life Sciences, University of Applied Sciences, FH Campus Wien.
authors.forename: Judit
authors.initials: J
authors.lastname: Fazekas
authors.affiliation: Department of Immunology, Capital Medical University, Beijing, PR China.
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.affiliation: Comparative Immunology and Oncology, Institute of Pathophysiology and Allergy Research, Medical University of Vienna.|Comparative Medicine, Messerli Research Institute, University of Veterinary Medicine Vienna, Medical University Vienna and University Vienna.
authors.forename: Marlene
authors.initials: M
authors.lastname: Weichselbaumer
authors.affiliation: Comparative Immunology and Oncology, Institute of Pathophysiology and Allergy Research, Medical University of Vienna.
authors.forename: Miroslawa
authors.initials: M
authors.lastname: Matz
authors.affiliation: Department of Biotechnology, VIBT-BOKU, University of Natural Resources and Life Sciences, Vienna, Austria.
authors.forename: Alexander
authors.initials: A
authors.lastname: Mader
authors.affiliation: Department of Biotechnology, VIBT-BOKU, University of Natural Resources and Life Sciences, Vienna, Austria.
authors.forename: Willibald
authors.initials: W
authors.lastname: Steinfellner
authors.affiliation: Comparative Immunology and Oncology, Institute of Pathophysiology and Allergy Research, Medical University of Vienna.|Comparative Medicine, Messerli Research Institute, University of Veterinary Medicine Vienna, Medical University Vienna and University Vienna.
authors.forename: Sarah
authors.initials: S
authors.lastname: Meitz
authors.affiliation: Comparative Immunology and Oncology, Institute of Pathophysiology and Allergy Research, Medical University of Vienna.
authors.forename: Diana
authors.initials: D
authors.lastname: Mechtcheriakova
authors.affiliation: Comparative Immunology and Oncology, Institute of Pathophysiology and Allergy Research, Medical University of Vienna.
authors.forename: Yuri
authors.initials: Y
authors.lastname: Sobanov
authors.affiliation: Department for Companion Animals and Horses, University of Veterinary Medicine Vienna.
authors.forename: Michael
authors.initials: M
authors.lastname: Willmann
authors.affiliation: Centre for Physiology and Pharmacology, Medical University of Vienna.
authors.forename: Thomas
authors.initials: T
authors.lastname: Stockner
authors.affiliation: Institute of Biochemistry and Molecular Biology, University of Hamburg, Hamburg, Germany.
authors.forename: Edzard
authors.initials: E
authors.lastname: Spillner
authors.affiliation: Department of Biotechnology, VIBT-BOKU, University of Natural Resources and Life Sciences, Vienna, Austria.
authors.forename: Renate
authors.initials: R
authors.lastname: Kunert
authors.affiliation: Comparative Immunology and Oncology, Institute of Pathophysiology and Allergy Research, Medical University of Vienna.|Comparative Medicine, Messerli Research Institute, University of Veterinary Medicine Vienna, Medical University Vienna and University Vienna.
authors.forename: Erika
authors.initials: E
authors.lastname: Jensen-Jarolim
mesh_terms: D000818:Animals; D016466:CHO Cells; D048708:Cell Growth Processes / Q000276:immunology; D002942:Circular Dichroism; D006224:Cricetinae; D003412:Cricetulus; D004283:Dog Diseases / Q000276:immunology / Q000628:therapy*; D004285:Dogs; D066246:ErbB Receptors / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D007116:Immunization, Passive / Q000379:methods*; D007074:Immunoglobulin G / Q000276:immunology*; D008958:Models, Molecular; D009369:Neoplasms / Q000276:immunology / Q000378:metabolism / Q000628:therapy / Q000662:veterinary*; D014162:Transfection
chemical_list: D007074:Immunoglobulin G; D066246:ErbB Receptors
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