PROTEIN PRODUCTION PLATFORM (P-CUBE)

The University of Zurich  (UZH) provides access to users of its novel DARPin technology in the Binder Selection facility of the Biochemistry Department of UZH. Ankyrin repeat proteins are ubiquitous protein-protein interaction molecules and were used as template to design ankyrin repeat proteins with fixed framework residues and randomised surface residues suitable for target binding. The random assembly of such modules yields combinatorial libraries of native Designed Ankyrin Repeat Proteins (DARPins) of varying length and diversities larger than 10¹⁰. By using ribosome display, an in vitro selection technique, specific binders with nanomolar affinities for almost any target protein can be selected. This opens the possibility to crystallise a target protein in complex with a variety of DARPins and enhances the chances of obtaining structures of target proteins that are otherwise not amendable to crystallization. The usefulness of this technology has been demonstrated for a variety of different protein classes such as proteases, kinases and membrane proteins. The selected ankyrin binders can easily be produced and serve as chaperones for crystallisation, in biochemical analyses such as western blot analysis, ELISA etc. This technology is unique in Europe (and world-wide).

Modality of access and work

Selection of binders using ribosome display takes approximately 8 to 12 weeks including the analysis of binding affinities. Users must be present part of the time to carry out experiments under the supervision of experts. At the end of the project, the user will have a set of DARPin molecules, which can be utilised for further experiments in their home laboratory. A typical duration of stay of users is about 8 weeks.

Support offered

At UZH, libraries of DARPins are available and infrastructure and reagents to perform screening by ribosome display will be offered. The ankyrin selection procedure is performed by personnel from the infrastructure, analysis of the pool of binders obtained will be done by personnel of the applicant laboratory. UZH has experience in guiding visitors on site as well as in providing expertise for the experiments to be carried out in the visitor's laboratory.