Padualaan 8, 3584 CH Utrecht
Paul van Bergen en Henegouwen
Nanobodies or VHHs consist of the variable region of heavy-chain antibodies that are present in animals from the family of camelidae. The Utrecht Nanobody Facility (UNF) offers technology to select and produce nanobodies to any possible target, including proteins and lipids. Selection of nanobodies will be done using phage display.
Nanobodies can be used for both bio-imaging and medical imaging. For bio-imaging, we conjugate nanobodies directly to gold particles of any size, which can used in immune-electron microscopy. We conjugate them to fluorophores, to be used in fluorescence microscopy including single molecule imaging, FRET and superresolution microscopy. For medical imaging, we couple nanobodies to radioactive tracers for SPECT or PET and to near-infrared dyes to be used in optical molecular imaging. Nanobodies can also be used to target nanoparticles as liposomes or micelles to desired sites in the mouse such as tumor xenografts.
Nanobodies have recently proven their role in the elucidation of the crystal structure of plasmamembrane proteins. Binding of a nanobody can stabilize plasmamembrane proteins in a particular conformation, which can be beneficial for the formation of protein crystals.
- Biomedical & health
- Industrial biotech
- single molecule imaging
- immuno-electron microscopy
- (superresolution) light microscopy
- protein purification
- Structural Biology
Expertise and Track Record
The Utrecht Nanobody Facility offers both selection and production of nanobodies in combination with technology for their application in molecular imaging, which includes single molecule imaging, immune-EM, fluorescence microscopy, in vivo imaging. This will be performed in collaboration with the Biology Imaging Center (BIC) from the Utrecht University.
– In vivo imaging of growth factor receptor as EGFR and Her2.
(Oliveira et al. Mol. Imaging 11; 33 2012; Kijanka et al. Eur J Nucl Med Mol Imaging. 40:1718 (2013).
– Targeting of liposomes, micelles en nanoparticles to cancer cells.
(Oliveira et al. J Con. Rel 145: 145 (2010); Talleli et al Journal of Control. Release 151:183 (2011); Heukers et al. Biomaterials 35;601 (2014)).
– Single molecule imaging of EGFR in plasma membrane of life cells.
(Low-nam et al. Nat Struct Mol Biol. 18:1244 (2011).
– Co-localization studies using Forster resonance energy transfer (FRET).
(Hofman et al. J. Cell Sci. 121: 2519 (2008)).
– Targeting of photosensitizers to cancer cells to perform photo dynamic therapy (PDT).
(Heukers et al. Nanomedicine 35: 601 (2014)).
I have experience in several collaborations aimed at the application of nanobodies in cancer diagnosis (Wegner et al.), cancer imaging (Vosjan et al., Van Driel et al) and therapy (Zeiss et al., Van Driel et al., Van der Water et al., Altintas et al., Van Meel et al.)
- van Driel PB, van der Vorst JR, Verbeek FP, Oliveira S, Snoeks TJ, Keereweer S, Chan B, Boonstra MC, Frangioni JV, van Bergen En Henegouwen PMP, Vahrmeijer AL, Lowik CW. 2014 Intraoperative fluorescence delineation of head and neck cancer with a fluorescent Anti-epidermal growth factor receptor nanobody. Int J Cancer. 134(11): 2663-2673.
- Wegner KD, Lindén S, Jin Z, Jennings TL, Khoulati RE, van Bergen En Henegouwen PMP, Hildebrandt N. Nanobodies and Nanocrystals: Highly Sensitive Quantum Dot-Based Homogeneous FRET Immunoassay for Serum-Based EGFR Detection. Small.
- 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 21:1214-24.
- Zaiss DM, van Loosdregt J, Gorlani A, Bekker CP, Gröne A, Sibilia M, van Bergen en Henegouwen PMP, Roovers RC, Coffer PJ, Sijts AJ. Amphiregulin enhances regulatory T cell-suppressive function via the epidermal growth factor receptor. Immunity. 2013 Feb 21;38(2):275-84.
- Van de Water JA, Bagci-Onder T, Agarwal AS, Wakimoto H, Roovers RC, Zhu Y, Kasmieh R, Bhere D, Van Bergen en Henegouwen PMP, Shah K. Therapeutic stem cells expressing variants of EGFR-specific nanobodies have antitumor effects. Proc Natl Acad Sci U S A. 2012 Oct 9;109(41):16642-7.
- Vosjan, M.W.J.D., Perk, L.R., Roovers, R.C., Visser, G.W.M., Stigter-van Walsum, M., van Bergen en Henegouwen, P.M.P. and van Dongen, G.A.M.S. . (2011) Facile labeling of an anti-epidermal growth factor receptor Nanobody with Ga-68 via a novel bifunctional desferal chelate for immuno-PET Eur J Nucl Med Mol Imaging. 38: 183-192.
- Van der Meel R, Oliveira S, Altintas I, Haselberg R, van der Veeken J, Roovers RC, van Bergen En Henegouwen PM, Storm G, Hennink WE, Schiffelers RM, Kok RJ. (2012) Tumor-targeted Nanobullets: Anti-EGFR nanobody-liposomes loaded with anti-IGF-1R kinase inhibitor for cancer treatment. J Control Release. 159:281-289.
- 1 postdoc
- 2 technicians
- 1 lab manager
- Standard molecular biology equipment including savety work benches, centrifuges and Akta FPLC systems