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

Sci Rep. 2024 Feb 14;14(1):3689. doi: 10.1038/s41598-024-53386-5.

Abstract

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.

MeSH terms

  • Animals
  • Antibodies, Monoclonal
  • Enzyme-Linked Immunosorbent Assay
  • Mammals / metabolism
  • Peptide Library
  • Phosphoric Monoester Hydrolases / metabolism
  • Single-Chain Antibodies*
  • Struthioniformes* / metabolism

Substances

  • Single-Chain Antibodies
  • Peptide Library
  • Phosphoric Monoester Hydrolases
  • Antibodies, Monoclonal