The development of canine immunotolerant monoclonal antibodies can accelerate the invention of new medicines for both canine and human diseases. We develop a methodology to clone the naive, somatically mutated variable domain repertoire from canine B cell mRNA using 5'RACE PCR. A set of degenerate primers were then designed and used to clone variable domain genes into archival "holding" plasmid libraries. These archived variable domain genes were then combinatorially ligated to produce a scFv M13 phage library. Next-generation long-read and short-read DNA sequencing methodologies were developed to annotate features of the cloned library including CDR diversity and IGHV/IGKV/IGLV subfamily distribution. A synthetic immunoglobulin G was developed from this scFv library to the canine immune checkpoint receptor PD-1. This synthetic platform can be used to clone and annotate archived antibody variable domain genes for use in perpetuity in order to develop improved preclinical models for the treatment of complex human diseases.

Faculty of Health Sciences Department Applied Biomedical Science Centre for Molecular Medicine and Biobanking University of Malta Msida Malta

Inserm UMRS1131 Institut de Génétique Moléculaire Université Paris 7 Hôpital St Louis Paris France

International Centre for Cancer Vaccine Science University of Gdansk Gdansk Poland

International Centre for Cancer Vaccine Science University of Gdansk Gdansk Poland; University of Edinburgh Institute of Genetics and Cancer Edinburgh Scotland UK

Research Centre for Applied Molecular Oncology Masaryk Memorial Cancer Institute Brno Czech Republic

Research Centre for Applied Molecular Oncology Masaryk Memorial Cancer Institute Brno Czech Republic; Laboratory of Growth Regulators Institute of Experimental Botany The Czech Academy of Sciences Olomouc Czech Republic

The University of Edinburgh Royal School of Veterinary Studies and Roslin Institute Edinburgh UK

University of Aberdeen Polwarth Building Foresterhill Aberdeen UK

University of Edinburgh Institute of Genetics and Cancer Edinburgh Scotland UK

University of Edinburgh Institute of Genetics and Cancer Edinburgh Scotland UK; Inserm UMRS1131 Institut de Génétique Moléculaire Université Paris 7 Hôpital St Louis Paris France

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