Deuteration and Macromolecular Crystallisation (DEMAX)

The three team members in DEMAX - Hanna Wacklin-Knecht, Oliver Bogojevic & Anna Leung (LTR)

DEMAX team members working on SINE2020 work package 5: (LtR) Hanna Wacklin-Knecht, Oliver Bogojevic & Anna Leung

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

The DEMAX platform produces deuterated materials such as biomass, proteins, lipids and other small molecules for neutron techniques such as small angle scattering, reflectometry, protein crystallography, spectroscopy, and powder diffraction. DEMAX also supports crystallization optimization for large crystal growth (protiated and/or perdeuterated) of protein for crystallography. This support functionality is particularly relevant for researchers in the fields of life science, soft condensed matter, chemistry, and biomaterials.

DEMAX entered initial operations in 2019 with the first pilot call for deuteration and crystallization proposals. During 2019-2021 DEMAX will run three of these pilot calls. The goals is to establish our procedures, complete fit-out of the lab, develop our proposal system, and practice serving users to be ready for hot commissioning and support for first science at ESS.

Scientists can submit proposals for deuterated materials for use in their neutron experiments at other facilities. Access is based upon peer review of proposals and scientific merit and is currently not limited to ESS member countries. The next call for proposals will be in late 2019 and will be advertised at https://deuteration.net. For more information email demax@esss.se.

Life Science & Soft Condensed Matter

ESS will enable some areas of life science, medical and pharmacological research to investigate with neutrons for the first time.

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The DEMAX platform consists of a chemistry laboratory and a biology/crystallisation laboratory. Our chemistry lab is shared with other ESS stakeholders and is leased from Medicon Village. The biology and crystallisation lab is co-housed with the Lund Protein Production Platform, LP3, within the biology department of Lund University.

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 (yeast, certain algal strains, bacteria), protein purification using liquid chromatography, some characterisation (DLS, SDS-PAGE, UV/Vis, thermal stability) and deuterium incorporation if feasible. 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 ionisation, GC-mass spec, and NMR).

Small angle 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

Chemical deuteration of small molecules can be achieved through a range of methods, and the modification of deuterated molecules using synthetic chemistry techniques allows a broad range of deuterated molecules to be synthesised. The ESS chemical deuteration laboratory is equipped to produce and modify a range of deuterated small molecules including lipids, surfactants and monomers.

ESS also focuses on the development of new techniques for deuteration chemistry, including enzyme-catalysed synthesis of challenging small molecules, and the chemical or enzymatic modification of biologically-produced materials.

Crystallisation

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 and often 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 maximise 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 and macrobatch support, large volume vapor diffusion set-ups, and a variety of temperature-controlled incubators for temperature-ramping experiments.

  • Establish procedures and protocols for deuteration and crystallisation for the user programme.
  • Enable a custom range of deuteration of small molecules material for use in small-angle neutron scattering, neutron reflectometry, and neutron diffraction (SINE2020-WP5).
  • Develop comprehensive support for large crystal growth under a range of different volumes and conditions (SINE2020-WP6 and Bergen in-kind). 

The ESS DEMAX platform exists as part of a broader network of deuteration laboratories within Europe and around the world (the deuteration network, DEUNET). The DEUNET aims to 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.

DEMAX is involved in two work packages 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.

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 maximise expertise and resources. DEMAX user labs are co-hosted with the LP3 at Lund University.

DEMAX is also involved in BrightnESS2, an EU-funded project within the European Commission’s Horizon 2020 research and innovation program: the ESS chemical deuteration laboratory has partnered with the deuteration facility at STFC to produce novel deuterated lipids for neutron experiments.

DEMAX also participates in LENS (League of Advanced European Neutron Sources), a consortium formed to promote cooperation between European neutron infrastructures offering transnational user programmes (working group 3: deuteration technologies).

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

Zoe Fisher

Group Leader for Deuteration and Macromolecular Crystallisation

ESS

Hanna Wacklin

Instrument Scientist - Neutron Reflectrometry

ESS

Anna Leung

Deuteration Scientist

ESS

Oliver Bogojevic

Junior Engineer

ESS