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Canine CD117-Specific Antibodies with Diverse Binding Properties Isolated from a Phage Display Library Using Cell-Based Biopanning

1
Australian Institute for Bioengineering and Nanotechnology (AIBN), The University of Queensland, Brisbane, QLD 4072, Australia
2
Faculty of Pharmacy, King Abdulaziz University, Jeddah 21589, Saudi Arabia
3
Vaccines and Immunotherapy Unit, King Fahd Medical Research Center (KFMRC), King Abdulaziz University, Jeddah 21589, Saudi Arabia
4
Australian Research Council Training Centre for Biopharmaceutical Innovation, The University of Queensland, Brisbane, QLD 4072, Australia
5
Centre for Advanced Imaging, The University of Queensland, Brisbane, QLD 4072, Australia
6
School of Veterinary Science, The University of Queensland, Gatton, QLD 4343, Australia
*
Author to whom correspondence should be addressed.
Antibodies 2019, 8(1), 15; https://doi.org/10.3390/antib8010015
Received: 11 November 2018 / Revised: 28 December 2018 / Accepted: 29 January 2019 / Published: 12 February 2019
(This article belongs to the Special Issue Antibody Phage Display)
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PDF [4551 KB, uploaded 12 February 2019]
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Abstract

CD117 (c-Kit) is a tyrosine kinase receptor that is overexpressed in multiple dog tumors. There is 100% homology between the juxtamembrane domain of human and canine CD117, and many cancer-causing mutations occur in this region in both species. Thus, CD117 is an important target for cancer treatment in dogs and for comparative oncology studies. Currently, there is no monoclonal antibody (mAb) specifically designed to target the exposed region of canine CD117, although there exist some with species cross-reactivity. We panned a naïve phage display library to isolate antibodies against recombinant CD117 on whole cells. Several mAbs were isolated and were shown to bind recombinant canine CD117 at low- to sub-nanomolar affinity. Additionally, binding to native canine CD117 was confirmed by immunohistochemistry and by flow cytometry. Competitive binding assays also identified mAbs that competed with the CD117 receptor-specific ligand, the stem cell factor (SCF). These results show the ability of our cell-based biopanning strategy to isolate a panel of antibodies that have varied characteristics when used in different binding assays. These in vitro/ex vivo assessments suggest that some of the isolated mAbs might be promising candidates for targeting overexpressed CD117 in canine cancers for different useful applications. View Full-Text
Keywords: Affinity selection; cancer; canine; CD117; c-Kit; dog; immunohistochemistry; membrane proteins; monoclonal antibody; phage display; sphere culture; transfection; veterinary oncology; whole cell panning Affinity selection; cancer; canine; CD117; c-Kit; dog; immunohistochemistry; membrane proteins; monoclonal antibody; phage display; sphere culture; transfection; veterinary oncology; whole cell panning
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Alfaleh, M.A.; Arora, N.; Yeh, M.; de Bakker, C.J.; Howard, C.B.; Macpherson, P.; Allavena, R.E.; Chen, X.; Harkness, L.; Mahler, S.M.; Jones, M.L. Canine CD117-Specific Antibodies with Diverse Binding Properties Isolated from a Phage Display Library Using Cell-Based Biopanning. Antibodies 2019, 8, 15.

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