• BIOCRYSTAL FACILITY

    AT
    THE CNR INSTITUTE OF BIOLOGY AND MOLECULAR PATHOLOGY
    AND
    THE DEPARTMENT OF BIOCHEMICAL SCIENCES, UNIVERSITY “LA SAPIENZA”

    Download Biocrystal Facility in pdf

PURPOSES

The Importance of Structural Biology

Structural biology is a powerful field that combines biophysics, biochemistry, molecular biology, and bioinformatics. It has fundamental applications in both biomedicine and biotechnology.

Macromolecular Crystallography: A Key Technique

Macromolecular crystallography is a critical technique used in structural biology. Advancements in synchrotron radiation sources, protein expression, and automated protein crystal growth have made it more accessible to molecular biologists.

Challenges and Opportunities for Italian Researchers

Despite these advancements, Italian researchers don’t fully utilize access to these technologies and large international facilities. This is particularly concerning in light of future European programs that emphasize large-scale research infrastructure.

Bridging the Gap: The Biocrystal Facility

To address this gap, the Biocrystal Facility connects Italian researchers with large international facilities, increasing their use of these resources. The facility assists users throughout the structure determination process, from construct selection and protein production to crystallization, data collection, and analysis.

ITACA.SB Network and Instruct Eric

Since November 2022, the Biocrystal Facility has been part of the ITACA.SB network. ITACA.SB aims to strengthen Italian capacity for structural biology services within the “Potentiating the ITAlian CApacity for Structural Biology Services in Instruct Eric (ITACA.SB)” project, funded by the European Union NextGenerationEU. Instruct Eric is a pan-European initiative that provides researchers worldwide with access to cutting-edge structural biology techniques and services.

The Biocrystal Facility: A Resource for Italian and International Researchers

The Biocrystal Facility, established through a collaborative effort between Sapienza University of Rome and the Italian National Research Council’s Institute of Molecular Biology and Pathology, offers Italian and international researchers access to crystal production and structure determination for biological macromolecules.

Resources

State-of-the-Art Equipment

Thanks to funding from the Sapienza University of Rome’s Equipment Grant 2020 and the ITACA.SB network, the Biocrystal Facility boasts cutting-edge equipment for high-throughput crystallization of both recombinant soluble and membrane proteins.

  • Oryx4 crystallization robot (Douglas Instruments UK) with an LCP module for protein crystallization (sitting/hanging drops), microbatch screening, optimization, and microseeding.
  • Oryx8 crystallization robot (Douglas Instruments UK) with an LCP module for protein crystallization (sitting/hanging drops), microseeding, and multivariate optimization (central composite and box-behnken).
  • Eppendorf epmotion 96 liquid handler for precise semi-automated pipetting.
  • Two Zeiss SteREO Discovery V20 KMAT stereomicroscopes offering magnification up to 96.5x and 150x. LED light sources provide clear field, dark field, and oblique illumination. The microscopes can be used with polarization lenses to identify crystals by birefringence. One microscope has a color Axiocam208 camera (4K resolution, 30 fps, 8.3 megapixels) for capturing images and videos.
  • Thermofisher Scientific NanoDrop One for rapid protein concentration determination using small volumes.
  • Temperature-controlled cabinets at 21°C, 4°C, and 37°C with vibration-proof motors.
  • TG40 crystallization condition temperature tester (Centeo) for simultaneously testing 5 different temperatures (2°C to 60°C) in a 40-well miniscreen.
  • Commercial crystallization screens:
    • Hampton Research: Index, Crystal Screen, and Stock Option
    • Qiagen: AmSo4, ProComplex, MPD, JCSG Core III & IV, Nucleix
    • Molecular Dimensions: Proplex, LMB, Morpheus, and MPD
    • Jena Biosciences: Wizard, JCSG++, PACT++, LCP
  • Coming Soon: XtalLight 100 (Xtal Concepts) for identifying protein crystals from salt crystals using intrinsic tryptophan fluorescence.

Beamline Access

In addition to its in-house equipment, the Biocrystal Facility offers access to beamlines at the European Synchrotron Radiation Facility (ESRF) in Grenoble, France, and ELETTRA Sincrotrone Trieste in Italy.

ACCESS PROCEDURES

The current staff capacity limits the number of external requests that can be accommodated. To submit an access request, please use the following link: https://www.cerm.unifi.it/instruct-it/submit-a-proposal

Click on Instruct Italia request Access

Select centers → Access to Biocrystal Facility → High Throughput crystallization or Crystals Optimization

Proposal Evaluation

The Scientific Council, composed of Andrea Ilari (ITACA.SB unit manager), Adele Di Matteo (crystallization expert), and Cécile Exertier (technical manager), will evaluate the feasibility of submitted proposals.

BIOCRYSTAL FACILITY ACKNOWLEDGEMENTS

Please acknowledge funding and resources as follows:

“The authors acknowledge the Biocrystal Facility and the support from the project ‘Potentiating the Italian Capacity for Structural Biology Services in Instruct Eric (ITACA.SB)’ (Project No. IR0000009) within the call MUR 3264/2021 PNRR M4/C2/L3.1.1, funded by the European Union NextGenerationEU.”

Knowledge Dissemination

The Biocrystal Facility actively disseminates knowledge related to biocrystallography and its services. This includes presentations at CNR conferences, information on the CNR website, targeted communications to potentially interested users, and courses highlighting the potential of biocrystallography for specific research projects.

STAFF

A Scientific Committee will verify the adequacy of the proposals submitted by users. It will consist of a structural biologist, a molecular biologist  and the Head of Service.

People
Activities shall be conducted by the following people, coordinated by Dr. Andrea Ilari.

Dr.
ANDREA ILARI

CNR – IBPM

Prof.
BEATRICE VALLONE

Sapienza UniRoma – CNR IBPM

Dr.
CARMELINDA SAVINO

CNR – IBPM

Dr.
ADELE DI MATTEO

CNR – IBPM

Dr.
GIANNI COLOTTI

CNR – IBPM

Giorgio Giardina Sapienza UniRoma

Prof.
GIORGIO GIARDINA

Sapienza UniRoma

Annarita Fiorillo Sapienza UniRoma

Prof.
ANNARITA FIORILLO

Sapienza UniRoma

Linda Celeste Montemiglio Sapienza UniRoma

Dr.
LINDA CELESTE MONTEMIGLIO

CNR IBPM

Dr.
CECILE EXERTIER

CNR IBPM

Dr.
FRANCESCA TROILO

CNR IBPM

Dr.
VERONICA MOREA

CNR – IBPM

STRUCTURE OF THE FACILITY

The facility is based on practical and theoretical consultancy at critical steps of protein crystallography, and it is also devoted to dissemination and training.
  1. Setting the biomedical or biotechnological project in the context of structural biology: defining objectives and critical points.
  2. Consulting in the production and purification of proteins.
  3. Crystallization screening using robot, their analysis and optimization initially with our in-house equipment and then supporting the application to European initiatives.
  4. Collection of Diffraction data at synchrotron radiation (ESRF).
  5. Determination of the three-dimensional structure and its analysis (in scientific collaboration).
  6. Assistance and training, in collaboration with ESRF and with groups of molecular biology, for the determination and analysis of the three-dimensional structure of proteins and their functional complexes.

WOW TO APPLY

Access is simple and informal. It is divided into two phases. The first will have a direct contact for guidance with one of the senior members of the Service (via email, phone or skype). In the second the use will fill in a form with a brief description of the project, which will be evaluated by the Scientific Committee. The accepted projects will follow the procedure described on page STRUCTURE OF THE FACILITY Each user will be assigned a scientific manager who will prepare the feasibility study and coordinate the operations of crystallization and data collection. The Facility will interact with users through a personal page on the website, on which materials related to a single project will be available, such as results of bioinformatics, images of crystallization trials, diffraction data and results of analysis of the stability and mono-dispersion of protein samples. The preferred mode for regular updates will be video conferences on the progress of each crystallization project.

USERS AND COLLABORATIONS

Interactions with other projects of CNR, including PON and POR, and with groups from the University “La Sapienza” will be favored. Other Universities and “not for profit” Institutions may be entitled to access, with the participation to the costs of each project. Access to payment for other types of users will require specific agreements, especially with respect to access to synchrotron facilities (ESRF).

EVALUATION OF THE PERFORMANCE OF THE FACILITY

The activities of the Service will be reviewed annually by the Scientific Committee.

CONTACT

Inviaci email

This contact form is deactivated because you refused to accept Google reCaptcha service which is necessary to validate any messages sent by the form.

How to reach us

The Department is located in the main campus of the University.
The main entrance of the University is in P.le Aldo Moro, 5 however the secondary entrance on Viale Regina Elena, 332 is closer to the Department.

By Train
The Department is about 1.5 Km from the main railway station (Termini). From there, you can either take a taxi (please use only official white taxis) or take the metro line B direction Rebibbia, exit at Policlinico Station (2nd stop) and walk for 500m to the Department. If you like, you can also walk here.

Fiumicino Airport
Take the fast train Leonardo Express to Termini. It leaves every 30 mins (at :07 and :37) starting from 6:37 until 23:37. The ticket costs 14.00 Euro one way.

Ciampino Airport
There are two bus services to Termini: Terravision and SITBus. Tickets cost 8 Eur one way and Eur 12.50 return (there are cheaper options if you buy them on the web). The bus leaves in connection with incoming flights and the ticket boot is in front of the Arrival gate. Depending on traffic, it might take between 30 and 50 mins.
You can also take a taxi. During the day the cost should be around 35-45 Euro, depending on traffic. Please only use official white taxis.