Utrecht Nanobody Facility

Mission

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.

Technology

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.

Applications

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.

References

Additional literature and examples of successful application of nanobodies:

Muyldermans S. Nanobodies: natural single-domain antibodies. Annu Rev Biochem. 2013;82:775-97. doi: 10.1146/annurev-biochem-063011-092449. Review.

Oliveira S, Heukers R, Sornkom J, Kok RJ, van Bergen En Henegouwen PM. Targeting tumors with nanobodies for cancer imaging and therapy. J Control Release. 2013 Dec 28;172(3):607-17. doi: 10.1016/j.jconrel.2013.08.298. Review.

Low-Nam ST, Lidke KA, Cutler PJ, Roovers RC, van Bergen en Henegouwen PM, Wilson BS, Lidke DS. ErbB1 dimerization is promoted by domain co-confinement and stabilized by ligand binding. Nat Struct Mol Biol. 2011 Oct 23;18(11):1244-9.
doi: 10.1038/nsmb.2135.

Oliveira S, van Dongen GA, Stigter-van Walsum M, Roovers RC, Stam JC, Mali W, van Diest PJ, van Bergen en Henegouwen PMP. Rapid visualization of human tumor xenografts through optical imaging with a near-infrared fluorescent anti-epidermal growth factor receptor nanobody. Mol Imaging. 2012 Feb;11(1):33-46.

Schmitz KR, Bagchi A, Roovers RC, van Bergen en Henegouwen PM, Ferguson KM. Structural evaluation of EGFR inhibition mechanisms for nanobodies/VHH domains. Structure. 2013 Jul 2;21(7):1214-24. doi: 10.1016/j.str.2013.05.008.

Nanobodies and nanocrystals: highly sensitive quantum dot-based homogeneous FRET immunoassay for serum-based EGFR detection. Wegner KD, Lindén S, Jin Z, Jennings TL, el Khoulati R, van Bergen en Henegouwen PM, Hildebrandt N. Small. 2014 Feb 26;10(4):734-40. doi: 10.1002/smll.201302383.