The Utrecht Nanobody Facility (UNF) offers technology for the selection and production of nanobodies for academic and non-academic researchers. We offer technology for the functionalization of nanobodies using different, site specific conjugation methods of fluorophores (Alexa, Atto, NIR dyes etc.), drugs, gold particles etc. Functionalized nanobodies are excellent tracers for imaging purposes and in collaboration with the Biology Imaging Center we provide for single molecule imaging, super-resolution light microscopy, electron microscopy and in vivo molecular imaging.
Nanobodies are small antibody fragments (15 kDa) derived from heavy chain antibodies from animals from the camelidae family such as the llama. Nanobodies can be selected from (custom built) immune libraries using phage display. Extensive equipment is available for the thorough characterization of the nanobodies. Important parameters are affinity, specificity both in vitro and in vivo and production yield. Nanobodies can be produced at small scale and equipment is available for the large scale production both from E. coli and HEK cells.
Nanobodies can be used for different applications such as stabilization of protein conformation for crystallization, protein or vesicle purification (immunoprecipitation), imaging (both light- and electron microscopy), as biosensors and for therapeutical applications, for instance as anti-cancer drugs.
- Production of immune libraries.
- Selection of nanobodies that bind with high specificity and affinity to a proteinaceous target (usually <10 nM).
- Production and purification at different yield using E. coli or mammalian HEK cells.
- Fusion to different tags such as myc, his, EPEA, FLAG, GFP, RFP etc.
- Site directed conjugation to fluorophores (Alexa, Atto, NIR dyes) or gold particles.
- Nanobodies can be engineered in different formats including bivalent, bispecific, trimeric, biparatopic etc.
- Nanobodies can be N-terminally biotinylated.
Typical time lines are three months for immunization and library construction and 3 months for selection.