Tiny tools closing the gap: Nanobodies in research and therapy

Nanobodies, also known as single-domain antibodies, have become powerful tools in research, diagnosis and therapy. Sourced from the heavy chain of camelid Heavy chain-only antibodies, this domain retains many important characteristics of full-length antibodies while being ~10 times smaller in molecular weight. Nanobody discovery has seen expansive development over the recent past, via both conventional antigen-exposed and completely synthetic repertoires. Along these lines, binding properties of candidates can be evolved by subsequent mutation and selection cycles to adjust their specificity and avidity. Due to their small size and compact structure, nanobodies can reach cryptic sites not accessible to conventional antibodies and also show superior tissue penetration. This penetrance, alongside their ease of handling, has made nanobodies ideal candidates for a myriad of immunotherapeutic and drug delivery applications. Furthermore, their small size imparts minimal linkage errors when conjugated to a fluorophore, making nanobodies ideal tools for high resolution imaging techniques. Most importantly, nanobodies can be stably expressed in living cells to bind, block or modify intracellular targets, enabling study of proteins in a native context at unprecedented detail. In this Review, we present the latest developments in nanobody technology and discuss applications in bioimaging, therapy, and intracellular protein study.