PROTEIN PRODUCTION PLATFORM (P-CUBE)

Transnational access to high-throughput crystallisation (HT-X) facilities will be coordinated by the EMBL Outstation at EMBL Hamburg (HAM) and provided by HT-X HAM and HT-X Grenoble (GRB) which have been fully operational for over 4 and 3 years, respectively. The crystallisation facilities at the University of Oxford (UOXF), Zurich (UZH) and EMBL Heidelberg (HD) are complementary installations that are unique to their location within this network.

HT-X Hamburg

The HT-X HAM is at least equal in size and capacity to Europe's largest installations. A distinguishing feature of the installation is its open and unrestricted access policy which had been implemented during its design phase and is reflected in its high degree of integration and automation. As a consequence, this installation has a proven track record in providing crystallisation services for a general user community for over 2 years. The automated crystallisation platform consists of two highly integrated modules: a liquid-handling station for the production of around 100 96-well crystallisation plates per day and a storage and imaging system with a capacity of 10,000 crystallisation plates (around 1,000,000 experiments). Crystallisation solutions are produced in-house and used for the automated set up of vapor diffusion (VD) experiments in 96-well crystallisation plates, as free-interface-diffusion (FID) experiments in micro-fluidic chips or as counter-diffusion (CD) experiments in capillaries. VD and FID experiments are automatically imaged and archived and the results are made available via internet and as DVDs, respectively. Capillaries are shipped to users for personal use and inspection.

HT-X Grenoble

One of the key platforms of the Partnership for Structural Biology (PSB) is the EMBL-GRN, which is located next to the European Synchrotron Radiation Facility (ESRF) and the Institute Laue Langevin (ILL). This facility offers completely automated crystallization screening in hanging and sitting drop set up at two temperatures ( 20 deg and 5 deg ). The facility consists of a high-volume liquid handling station for the production of screens, a Cartesian nanovolume crystallization robot and three automated imaging systems, two operating at 20 deg and one at 5 deg. Results together with all experimental parameters are made available to users in real time through our CRystallization Information Management System (CRIMS), which includes fast and convenient web user interfaces providing advanced tools like Drop Scoring, Drop history, Navigation through project targets and samples and project summaries among others. CRIMS is synchronized with the ESRF automated sample changers and data collection software through ISPyB. Crystallization plates containing crystals are made available to users upon requests (typically during their next synchrotron visit) and facilities for crystal mounting and freezing are provided. In the six years of operation this facility has provided crystallization screening to more than 300 scientists.

HT-X University of Oxford

The HT-X UOXF is similar to the HT-X HAM, but also offers crystallisations at 5°C. Crystallisation images are accessed through a web-interface.

HT-X University of Zurich

The HT-X UZH was installed in 2001 through the Swiss Consortium in Structural Biology which has a focus on structure determination of membrane protein and supra-molecular assembly structures by X-ray crystallography. In its 6th year in operation, the UZH HT-crystallisation facility serves 20 research groups with over 100 researchers from the UZH, the ETHZ, the UniBas and the PSI/SLS with initial- and refinement VD sitting-drop crystallisation screens and screens routinely up to 10,000 crystallisation conditions per day. For the VD crystallisation experiments in 3x96-well crystallisation plates (CrystalQuick; Greiner-BioOne), the HT-crystallisation facility offers 30 commercial and in-house screens for the initial screening. For refinement screens the HT-crystallisation facility routinely prepares custom-made and additive crystallisation screens tailored to the requirement of the protein. With its automation and capacity for processing crystallisation experiments, the installation is also accessible to any academic research groups as well as to small biotech companies. The HT-X UZH offers crystallisations at 4°C and 20°C.

All installations employ different equipment and methods that are complementary. Users gain access to a much more diverse set of techniques which will increase the chances for successful crystallisation.

Existing possibilities

Modality of access and work

HT-X HAM: Users of the HT-X HAM design and schedule the execution of their experiments on-line and have internet based access to the results. Samples are shipped to the facility (by mail or any other preferred mode) and set up according to the specifications set by the users.
Reservations for initial screening experiments are possible for the following working day. Optimisation experiments have to be booked 2 days in advance.

HT-X GRB

The user designs the experiments through our dedicated web interface. Details on the methods and possible experimental parameters are available through our web portal, however specific and advice can be requested from the HTX lab staff at This e-mail address is being protected from spambots. You need JavaScript enabled to view it . Every individual crystallization experiment is submitted to a quality control process. After the crystallization experiments are set up the user receives an experimental report including data on quality control. Results are available in real-time though our web-based Crystallization Information Management System (CRIMS), which is synchronized with the ESRF sample changer and data collection system ISPyB (this requires the use of SPINE sample loops with 2D-data matrix). Plates are made available to users upon requests, (typically during their next visit to the ESRF beam lines). If necessary, lenses and liquid nitrogen will be made available for crystal mounting and freezing (but no sample loops).

HT-X HD: experiment design and reservations are made through a common web page.

HT-X UZH: Users of the HT-X UZH have access through internet for scheduling their experiments and for viewing their plates-images at 4°C and 20°C. The "Image Now" function of the plate incubator and imaging system enables the researcher to access their plates through the internet in real time and assess the progress of the crystallisation experiment.

UOXF: UOXF provides scheduling of crystallization experiments on-line through xtalPIMS. This covers crystallization, i.e. creation of crystallization trials, monitoring of experiments through imaging systems, harvesting of crystals and managing data collection information and can be accessed via a web based GUI or an offline GUI which also provides imager integration.

Support offered

HT-X HAM: HT-X HAM has developed a recommended and prioritized set of initial experiments for users with less experience.HT-X HAM also offer an 'Expert Mode' for and during the design of optimisation experiments. The latter allows users to take advantage of the local know-how and expertise of the experienced team of crystallisers.

At HT-X GRB the laboratory staff is available for any specific request and can advice users on the best choices for their projects. In addition to the standard procedures, a number of alternative protocols are available to support samples with specific requirements (low stability, detergents etc,). Support for the design of optimisation experiments is also available. You can contact the laboratory staff at This e-mail address is being protected from spambots. You need JavaScript enabled to view it

HT-X UZH: For the past 6 years HT-X UZH has employed successfully an "assistant-on-demand" strategy. Users new to the field of protein crystallisation have unrestricted access to HT-X UZH expertise in crystallisation techniques.

HT-X UOXF: HT-X UOXF offers access to existing crystallisation technologies from the start of Pcube. It also will develop novel crystallisation technologies and these will then be offered for access in years 3 and 4. The new facilities which will become available are

• counter diffusion capillary crystallisation and
• in situ X-ray diffraction. The level of access will be in the order of 500 plates per year for crystallisation and X-ray diffraction. A small number of plates utilising these new technologies will be made available during years 1 and 2.