# Novel Type IIS-Based Library Assembly Technique for Developing Nanobodies Targeting IPNv VP2 Protein

## TL;DR

A new method for creating nanobodies targeting a virus affecting fish is developed, offering faster and more adaptable tools for disease control in aquaculture.

## Contribution

A novel Type IIS-based library assembly technique for nanobody development is introduced, enabling rapid and adaptable targeting of proteins.

## Key findings

- A nanobody targeting the VP2 protein of IPNv was successfully identified and validated.
- The method allows for the rapid generation of diverse nanobody repertoires adaptable to various protein targets.
- The pipeline integrates density gradient enrichment and high-throughput screening for efficient nanobody discovery.

## Abstract

The development of effective tools to combat viral diseases remains a major challenge for the aquaculture industry. Infectious pancreatic necrosis virus (IPNv) is one of the most devastating pathogens affecting salmonids, leading to high mortality rates and substantial economic losses worldwide. Here, we present a novel nanobody discovery pipeline based on a Type IIS restriction enzyme-driven library assembly method that enables the rapid generation of highly diverse nanobody repertoires. This streamlined approach not only shortens the time required for nanobody identification but also offers remarkable adaptability, allowing its application to virtually any protein target, including antigens from aquaculture pathogens and beyond. By integrating this strategy with density gradient–based enrichment and high-throughput screening, we successfully identified and validated a nanobody against the VP2 protein of IPNv, a key structural component essential for viral infectivity. These findings highlight the potential of this platform both as a versatile methodological advance in antibody engineering and as a practical foundation for developing innovative diagnostic and therapeutic tools. Ultimately, nanobodies generated through this pipeline could play a pivotal role in improving disease management and enhancing sustainability in aquaculture.