Integrating Computational Design, Synthetic Biology, and Delivery Platforms in Next-Generation Nanobody Technologies

Nanobody research has become a rapidly expanding field at the intersection of molecular biology, immunology, and bioengineering. Originally derived from camelid heavy-chain antibodies, nanobodies have demonstrated exceptional stability, affinity, and versatility compared to conventional antibodies, enabling their use across diagnostics, therapeutic interventions, imaging, and intracellular targeting. Over the past decade, major advances in synthetic library development, display technologies, and computational protein design have accelerated nanobody discovery and optimized their molecular properties. Moreover, AI-assisted structure prediction and binder screening are now reshaping how nanobody–antigen interactions are modelled and validated. Despite these promising developments, challenges remain in achieving precise intracellular delivery, ensuring long-term stability in complex in vivo environments, and expanding clinical translation through robust production systems and delivery routes. Addressing these knowledge gaps is crucial for transforming nanobody technologies into widely deployable biomedical solutions.

This Research Topic aims to gather multidisciplinary research and reviews that advance the full nanobody development pipeline—from target selection and binder discovery to molecular engineering, functional validation, and preclinical or clinical implementation. The goal is to showcase innovative strategies that enhance discovery throughput, increase binding specificity and stability, enable effective intracellular targeting, and translate nanobody platforms into therapeutic, diagnostic, and imaging applications. In doing so, this collection seeks to outline the state of the art while highlighting the technological and conceptual barriers that must be overcome to fully integrate nanobody-based systems into next-generation precision medicine.

To gather further insights into nanobody-based technologies and their biomedical applications, we welcome articles addressing, but not limited to, the following themes:

• Nanobody discovery technologies: synthetic, immune, or naïve libraries; phage, yeast, ribosome, bacterial, or mammalian display systems; AI-guided and computational screening; high-throughput sequencing; affinity maturation and engineering.

• Target identification and validation: discovery of nanobodies for intracellular and membrane-associated targets; tumor-associated antigens; neurodegenerative and infectious disease targets; multi-omics–driven target prioritization.

• Structural and functional engineering: humanization, multivalent and multi-specific nanobodies, solubility and stability optimization, intrabodies, protein–protein interaction modulation, structural biology of nanobody–target complexes.

• Nanobodies in diagnostics: biosensors, point-of-care and imaging technologies (PET/SPECT, live-cell, super-resolution microscopy), and diagnostic nanoprobes.

• Therapeutic and delivery platforms: nanobody-mediated drug delivery, AAV and CAR-based systems, gene therapy, targeted protein degradation, blood–brain barrier–penetrating nanobodies, RNA and CRISPR delivery.

• Translational and clinical applications: oncology, neurodegenerative and infectious diseases, autoimmune disorders, and precision medicine approaches.

Article types: Original Research, Review, Mini Review, Hypothesis and Theory, Perspective, and Methods.