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.

• Klíčová slova
• CP: cancer biology, CP: immunology, canine physiology, next-generation sequencing, scfv phage libraries, translational research, veterinary medicine,
• MeSH
• jednořetězcové protilátky * genetika   imunologie   MeSH
• lidé MeSH
• monoklonální protilátky MeSH
• nádory * imunologie   MeSH
• peptidová knihovna * MeSH
• psi MeSH
• rekombinantní proteiny izolace a purifikace   imunologie   genetika   MeSH
• translační biomedicínský výzkum * metody   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• psi MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• jednořetězcové protilátky * MeSH
• monoklonální protilátky MeSH
• peptidová knihovna * MeSH
• rekombinantní proteiny MeSH