Development of an ostrich-derived single-chain variable fragment (scFv) against PTPRN extracellular domain

• Published in: Scientific reports Vol. 14; no. 1; p. 3689
• Main Authors: Dabiri, Hamed, Sadeghizadeh, Majid, Ziaei, Vahab, Moghadasi, Zahra, Maham, Ali, Hajizadeh-Saffar, Ensiyeh, Habibi-Anbouhi, Mahdi
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 14-02-2024; Nature Publishing Group; Nature Portfolio
• Subjects: 631/1647/664/2228; 631/61/490; Animals; Antibodies, Monoclonal; Autoimmune diseases; Beta cells; Binders; Diabetes; Diabetes mellitus (insulin dependent); Enzyme-Linked Immunosorbent Assay; Flow cytometry; Homology; Humanities and Social Sciences; Immune system; Immunotherapy; Mammals; Mammals - metabolism; Monoclonal antibodies; multidisciplinary; Pancreas; Peptide Library; Phage display; Phosphoric Monoester Hydrolases - metabolism; Phylogeny; Protein-tyrosine-phosphatase; Science; Science (multidisciplinary); Single-Chain Antibodies; Struthioniformes - metabolism; Western blotting
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-024-53386-5

In type 1 diabetes, the immune system destroys pancreatic beta cells in an autoimmune condition. To overcome this disease, a specific monoclonal antibody that binds to pancreatic beta cells could be used for targeted immunotherapy. Protein tyrosine phosphatase receptor N (PTPRN) is one of the important surface antigen candidates. Due to its high sequence homology among mammals, so far, no single-chain monoclonal antibody has been produced against this receptor. In this study, we developed a novel single-chain variable fragment (scFv) against the PTPRN extracellular domain. To this aim, ostrich species was used as a host is far phylogenetically birds from mammals to construct a phage display library for the first time. An ostrich-derived scfv phage display library was prepared and biopanning steps were done to enrich and screen for isolating the best anti-PTPRN binders. An scFv with appropriate affinity and specificity to the PTPRN extracellular domain was selected and characterized by ELISA, western blotting, and flow cytometry. The anti-PTPRN scFv developed in this study could be introduced as an effective tool that can pave the way for the creation of antibody-based targeting systems in cooperation with the detection and therapy of type I diabetes.