The development of a canine single-chain phage antibody library to isolate recombinant antibodies for use in translational cancer research
Jazyk angličtina Země Spojené státy americké Médium print
Typ dokumentu časopisecké články
PubMed
40132540
DOI
10.1016/j.crmeth.2025.101008
PII: S2667-2375(25)00044-X
Knihovny.cz E-zdroje
- Klíčová slova
- CP: cancer biology, CP: immunology, canine physiology, next-generation sequencing, scfv phage libraries, translational research, veterinary medicine,
- MeSH
- antigeny CD279 imunologie MeSH
- jednořetězcové protilátky * imunologie genetika MeSH
- lidé MeSH
- monoklonální protilátky imunologie genetika MeSH
- nádory imunologie terapie MeSH
- peptidová knihovna MeSH
- psi MeSH
- rekombinantní proteiny imunologie genetika MeSH
- translační biomedicínský výzkum MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- psi MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- antigeny CD279 MeSH
- jednořetězcové protilátky * MeSH
- monoklonální protilátky MeSH
- peptidová knihovna MeSH
- rekombinantní proteiny MeSH
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.
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
Research Centre for Applied Molecular Oncology Masaryk Memorial Cancer Institute Brno 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
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