Deuteration and Macromolecular Crystallisation (DEMAX)

crystal transport tube

The Deuteration and Macromolecular Crystallisation (DEMAX) platform supports life science and soft matter research users of ESS.

This is done through support laboratories and facilities for biological deuteration of biomass, proteins, lipids and some small biomolecules. DEMAX also supports crystallization optimization for large crystal growth (hydrogenous or perdeuterated) macromolecules for neutron protein crystallography.

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Life Science & Soft Condensed Matter

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DEMAX is composed of laboratory facilities that support ESS users to help them achieve maximum scientific impact in the life sciences, soft condensed matter, and biomaterial scientific research.

ESS users will have access to a biodeuteration and macromolecular crystallography lab and service-level support for chemical deuteration. In this way users can access deuterated material production and large crystal growth for neutron experiments.

Biological Deuteration

Deuteration of biomolecules is achieved by culturing cells under deuterated conditions, harvesting the cell paste, and then processing or extracting the desired products. Deuterated products can include polysaccharides, proteins, lipids, and DNA/RNA. In some cases the extracted materials can be used for the derivation of other deuterated biomolecules. DEMAX will support cell paste production from a number of organisms, protein purification using liquid chromatography, and characterization (DLS, SDS-PAGE, UV/Vis, thermal stability). For other types of biomaterials, molecules are purified using basic chemical methods: solvent extraction, chromatographic purification (FPLC, HPLC) and their purity/composition is analysed using standard methods (such as gas chromatography (GC) with flame ionization, GC-mass spec, and NMR).

Small angle neutron scattering, spin echo, reflectometry, and protein crystallography are fast growing areas in soft matter and life science that have a need for these kinds of materials. For protein crystallography there is an enormous benefit to using deuterated proteins. Not only does deuteration boost the signal-to-noise ratio, it also allows for much smaller (~10 times) crystal volumes required. These labeled biomaterials can be fully, partially or selectively deuterated, as required for a particular experiment.

Chemical Deuteration

A node for molecular synthesis at ESS will be established to complement the existing deuterating capabilities for small organic molecules. This development will done in collaboration with the leading deuteration laboratories at ISISILL and FZJ as part of a chemical deuteration network (DEUNET). DEUNET will address the needs of the community by developing a cost-effective platform to provide access to a broad range of materials and expertise, new synthetic methods (products), synthesis of novel materials, and to coordinate service for European neutron users.

The ESS node (DEULAB) will focus on the chemical synthesis of complex deuterated molecules based on both biological and non-biological starting materials sourced from ISIS and ILL, and on developing novel deuteration methods. In particular, ESS will develop the enzyme-catalysed synthesis of chiral deuterated compounds, and synthetic transformations of biological molecules.

Crystallization

Large crystals (at least 0.1–0.5 mm3) are required for successful neutron protein crystallography experiments. Most crystals that are able to grow to 200-300 microns without explicit effort can be coaxed to larger sizes. This requires detailed knowledge of protein properties, the right approach, and tools. Using larger crystals offers additional advantages: higher quality data, higher resolution data, faster data collection times, and they are easier to manipulate. So, while ESS flux and beam characteristics will lower the required size, it is still advantageous to maximize the crystal volume. DEMAX will offer integrated support for protein production to large crystal growth. Included is access to an Opticrys dialysis crystallization device, Oryx8 screening robot, microbatch & macrobatch support, large volume vapor diffusion set-ups, and a variety of temperature-controlled incubators for temperature-ramping experiments.

We have laboratory spaces at Lund University dedicated to biodeuteration (set up for algae, bacteria and yeast), and macromolecular crystal growth.

In addition, we have chemistry laboratories dedicated to chemical deuteration. The latter are shared with the ESS Target Division, other Science Support Systems platforms and the ESS Neutron Instruments Division.

Development Activities

  • Establish procedures and protocols for bio-deuteration for the user program.
  • Enable a custom range of deuteration of some bio-derived small chemicals and biological material for use in small-angle neutron scattering, neutron reflectometry, and neutron protein chrystallography (SINE2020-WP5).
  • Develop comprehensive support for large crystal growth under a range of different volumes and conditions (SINE2020-WP6 and Bergen in-kind). 

Collaborations

DEMAX is involved in two work-pages in the EU-funded consortium SINE2020  (Science and Innovation with Neutrons in Europe in 2020): WP5 for chemical deuteration and WP6 for large crystal growth. This is in collaboration with ILL and MLZ.

Establish the DEUNET network, part of WP5. Coordinate efforts between multiple european deuteration labs.

The DEMAX platform is a part of a collaborative effort with the Lund Protein Production Platform (LP3), which is a cross-faculty Lund University resource. The collaboration serves to maximize expertise and resources. DEMAX user labs are co-hosted with the LP3 at Lund University.

In-Kind Partner

DEMAX has partnered with University of Bergen, Norway, to develop and partly equip the biodeuteration and crystallization labs shared with LP3. This collaboration has ensured that DEMAX will have liquid handling robots and incubators to support biodeuteration and large crystal growth.

Contact

DEMAX platform

Zoe Fisher

Support Scientist

ESS

Chemical deuteration

Hanna Wacklin

Instrument Scientist - Neutron Reflectrometry

ESS