DEMAX: update, current status

1. DEMAX: update, current status Dr. Zoë Fisher DEMAX platform lead Adjunct Snr. Lecturer at Lund University (Biology Dept) NMX STAP meeting2-3 October 2017

2. Outline • What is DEMAX? • Expanded scope since last NMX STAP • Staffing: current & future • Activities (DEUNET, SINE2020, Interreg) • Labs and location • Access & initial operations plan for '19–'23 • Conclusions

3. DEMAX is a user support lab for deuteration, crystallization • Advice, expertise, access to assist user with sample preparation,make the right sample, with the right amount of D incorporated for NPX, SANS, NR • DEMAX now includes: • Biological deuteration (proteins, lipid, or just the biomass) • Chemical deuteration (small molecules) • Large crystal growth

4. Current staffing • Zoë Fisher (sci) & 0.5 FTE (shared with LP3) *Biodeuteration (protein), macromolecular crystallization • Hanna Wacklin (sci) & Anna Leung (postdoc) *Chemical and biodeuteration (lipid, membranes, small molecules)

5. Planned staffing & ramp-up • In the future (2023) DEMAX will have 7 FTE, spread across the 3 pillars: • Biodeuteration: 1 FTE scientist, 1 FTE technician (possibility to share with LP3) • Crystallization: 1 FTE scientist, 1 FTE technician (possibility to share with LP3) • Chem. deuteration: 3 FTE scientists • Total ops budget: 1.225 M€/yr (includes: consumables, bench fees/rents, services, equipment – large, small, replacement, service). Ops FTE

6. Activities - biodeuteration • Develop protocols for protein expression (optimize yield, maximal D incorporation at lowest cost, characterize proteins made with labeled vs unlabeled C source..establish mass spec, biophysical characterization as part of work flow) • Shared FTE with LP3 is doing a lot of this work • MAX4ESSFUN-(Interreg)-funded projects (1 current project for protein deuterated for SANS..1 more application involving deuteration is planned)

7. Activities – chem deuteration • SINE2020 – enzyme-driven catalysis to produce labeled small molecules (Anna L.) • Establish DEUNET (European network for deuteration) • Small molecule synthesis, lipid extractions, purification, characterization • Postdoc starting on yeast lipid project Jan 2018 (VR funded) • Recruiting deuteration chemist at the moment (exp start date Q2 2018)

8. Activities – xtallography • SINE2020 – WP on large crystal growth, vapour diffusion & dialysis with temperature as variable • In-kind agreement with UiB (Norway) – Oryx8, Opticrys device, used in development activities for SINE2020 grant • Interreg projects (3 current projects for crystal growth, optimization; 2 more applications planned)

9. DEMAX & LU collaboration • To realize part of our mission we established a formal partnership with LU (spec. the LU Protein Production Platform, LP3) for biodeuteration, crystallization • ESS & Lund University signing a MoU (5 years). • DEMAX has a bench fee agreement with LP3 to enable formal partnering (5 years, started in 2016). • Includes lab space, office, and central services (milliQ, waste, etc) • For 24 months also share a technical FTE for "contract research" to develop and document protocols for DEMAX in biodeuteration strategies, benchmarking methods, take care of xtallography equipments/supplies/manuals etc.

10. Current location MAX IV ESS • Here we will specialize in deuterated biomass production (cell paste, bacteria & yeast & algae for lipids and/or protein extraction), deuterated protein purification (on request or through LP3), and large crystal growth (screening & optimization). DEMAXis now co-located with LP3 in Biology Department (LU) Bus service to ESS site (no. 20) – takes ~12 min, departs every 10 min. Replaced with tram in Fall 2019. KILU BMC Medicon Village Max lab

11. LP3 and DEMAX shared spaces in Biology Dept (MCB Unit) ** LP3 moved Dec 2015, DEMAX moved in July 2016

12. Chemical deuteration • Sharing space with Target/Science people in the ~100 m2 rented lab space in Medicon Village bldg 403. (lease ends Oct ’18 with possibility to extend – being negotiated) • Essential equipment is in place for synthesis, separation, characterization • For NMR we have arrangement with Red Glead, MS with LU Chemistry. MV Chemistry Lab 100 m2 ESS Headquarters

13. Current status of biodeuteration & crystallization • Protein expression, purification, basic characterization (purity, concentration) • Biophysical characterization (DLS, thermofluor, Nanotemper for temperature stability) • Crystallization hardware for screening (small & large volume: Moquito (LP3), Oryx8 (DEMAX), Opticrys (coming Q4 2017), Centeo (temp) • Tools & equipment for crystal mounting and H/D exchange for both RT and cryo X-ray/neutron data collection.

14. Initial operations 2019-2023 • Access policy for users to ESS is not in place • We intend to run the lab as a service (chem & biodeuteration, mixed for crystallography) • By 2019 we will have key staff in place (but not all) • We intend to make one call/year for "friendly user" proposals initially with an interim review panel (projects should be do-able and help us develop, while leaving enough resources to finish staffing, equipping labs before official start of User Program 2023) • When instruments are on-line, access will be tied to having awarded beam time and tracked in the same way

15. Conclusions • DEMAX is on-track to start initial "friendly users" ops in 2019 • Recruitment is underway for chemist, plan for technical recruit on bio/xtal side in Q1 2019 • Partnering with LU brings many opportunities, but also carries some long-term operational risk • Key issue for chemdeuteration is securing a suitable space beyond the MV lease.