Nanobody Generation Unit


Nanobody Generation Unit 1

The Nanobody generation unit is currently located in the Faculty of Science on the south-side of Brussels, in the city of Etterbeek. The unit is embedded within the lab 'Cellular and Molecular Immunology (CMIM)’ and is hosted by professors Serge Muyldermans, Patrick De Baetselier, Stefan Magez, Jo Van Ginderachter and Geert Raes. It also shares equipment and lab space with the Nanobody Service Facility headed by Dr. Ghlomareza Hassanzadeh Ghassabeh.

Nanobody Generation Unit 2

Nanobodies have been discovered two decades ago by a team at the Vrije Universiteit Brussel including Prof. Muyldermans. Over the years, his team and a consortium of collaborators further extended and improved the Nanobody knowledge and technology platform.
The pathway to the generation of nanobodies for their use in imaging applications can be diverse. It generally starts with the immunization of camelids with a source of the target antigen of choice, being most often a recombinant or purified protein, (transfected) cells or DNA encoding the target. Camelids (i.e. camel, dromedary, alpaca, vicuna or llama) are kept in farms and animalaria of a multitude of collaborators at various places, both in and outside of Belgium. At an optimal time-point, peripheral blood from immunized animals is collected and shipped to CMIM where it is processed to generate a so-called 'immune nanobody library'. As an alternative to immune libraries, also synthetic and naive nanobody libraries are at hand if immunization is cumbersome. In a next phase, individual antigen-specific nanobodies are isolated from these libraries. Although we have generated and have access to many nanobody selection techniques, we commonly use M13 phage display, for which we have dedicated a separate, isolated room with a hood, shakers and centrifuges. Once antigen-specific nanobodies are identified nanobodies can be further genetically manipulated in our dedicated molecular biology unit, where we have at hand multiple bacterial flows, PCR equipment, an agarose gel room, spectrophotometers, -80°C and -20°C freezers, cold rooms, water baths, incubators and a protein gel area. We also keep a large catalogue of all possible constructs as glycerol stock backup in our -80°C freezers.

Nanobody Generation Unit 3

The nanobody generation unit also has the possibility to produce and purify large quantities of nanobodies. Currently we are limited to recombinant E. coli cultures in shaker flasks, with a capacity of 20L culture per day. His-tagged nanobodies are commonly extracted and purified from bacterial cultures via osmotic shock, Nickel-NTA affinity chromatography and gel filtration. This can be performed semi-automatically on our dedicated AktaXpress system.

Nanobody Generation Unit 4

Purified nanobodies are extensively biochemically and biologically characterized within our unit. In-house analysis equipment include a fully equipped AktaExplorer System, ELISA and fluorescent plate readers, a nanodrop spectrophotometer and a T200 Biacore Surface Plasmon Resonance system. We also have 5 air flows and incubators for eukaryotic cell manipulations, an animalarium with a breeding facility and a capacity of about 200 cages, a FACS Canto flow cytometer and a FACSCalibur flow sorter with a specialized technician.

Contact:
Prof. Dr. Nick Devoogdt
Labo ICMI: 0032 (0)2 477 4991
Labo CMIM: 0032 (0)2 629 1978
ndevoogd@vub.ac.be