The discovery of heavy-chain-only antibodies (HCAbs) in camelids and sharks led to the rise of a new research field in which single-domain antibodies are used for various applications. Single-domain antibodies are the antigen-binding fragments derived from HCAbs showing several beneficial properties (e.g., small size, specificity, stability under extreme conditions, cost-effective production, and ease of engineering). Importantly, they are stable in reducing cytoplasmic environment, which allows their use as an intrabody to target a wide range of intracellular targets. In this chapter, we discuss both the therapeutic potential of camelid single-domain antibodies (nanobodies) and their use as a research tool with the main focus on its intracellular employment. Targeting intracellular proteins using nanobodies as a therapeutic per se is, up to now, limited due to its incapacity to traverse the cellular membrane. They can however serve as a stepping stone to small compound development, since they directly target a resident, endogenous protein, similar to how a conventional drug acts. In addition, nanobodies are highly adaptable tools and possess interesting properties for more fundamental research objectives like the elucidation of protein function, the tracking and visualization of endogenous proteins in an in vivo setting, and the assessment of protein-protein interactions.
Part of the book: Antibody Engineering