Identification and characterization of nanobodies specific for the human ubiquitin-like ISG15 protein.

ABSTRACT: Interferon-induced ubiquitin (Ub)-like modifier Interferon Stimulated Gene 15 (ISG15) functions both intracellularly and as a secreted protein with cytokine-like properties. The ISG15 pathway is implicated in various diseases, including cancer and inflammatory disorders, but understanding its precise roles has been challenging because of limited availability of tools to study ISG15 biology. Here, we report the development of two novel nanobodies that target human ISG15, obtained through alpaca immunization and phage display. These nanobodies, VHH_ISG15-A and VHH_ISG15-B, exhibit nanomolar binding affinities and recognize distinct epitopes on ISG15's C- and N-terminal domains, respectively, as demonstrated by NMR and X-ray structural analyses. Both nanobodies enable the

SUBMITTER: Gan J 

PROVIDER: S-EPMC12406269 | biostudies-literature | 2025 Sep

REPOSITORIES: biostudies-literature