Dr. Jeremy Keown, from the United Kingdom, collaborated with Instruct-ERIC and VIB VZW in Brussels through the ISIDORe project to generate nanobodies targeting viral polymerases from Bunyaviruses, which are responsible for several localized epidemics and could pose a pandemic threat in the future. The viral polymerase plays a crucial role in genome replication and transcription, but its flexibility makes it difficult to study using traditional structural methods.
The goal of the project was twofold: first, to use nanobodies to stabilize the flexible regions of the polymerase for better structural analysis; and second, to inhibit specific functions of the polymerase to track its location during viral infection.
The research team provided purified polymerase protein to the Nanobody Discovery facility at Instruct-BE, where it was injected into llamas. From this, 35 nanobodies were generated for one target, and 63 for a second polymerase target. These nanobodies are currently being expressed in E. coli for structural and in vitro functional experiments. Further work includes sub-cloning the nanobodies into mammalian expression plasmids to test their efficacy against viral infection and polymerase activity.
Once substantial progress has been made, the findings will be published in open-access journals.