Missions

Solid-state NMR spectroscopy has recently developed into an essential approach to investigate large supramolecular biological assemblies, which are difficult to crystallise and are too disordered for cryo-EM studies. The technique appears thus particularly adapted for the structure determination of adjuvant-attached antigens, which could potentially facilitate vaccine research. However, a number of fundamental developments still need to be made before the innovative potential of this new approach can be fully exploited.
The candidate will work on the development of new solid-state NMR strategies (particularly hyperpolarised) and the application of this spectroscopy to complex problems in the field of drugs and vaccines. The project will notably build on two recent additions in the solid-state NMR portfolio, namely dynamic nuclear polarisation (DNP) and 1H-detection at increased magic-angle spinning (MAS) frequencies, which address the intrinsic sensitivity problem of the technique and can be taken of advantage to perform structural studies of antigens in heterogenous vaccine formulations.
The project will involve: i) implementing new isotope labelling strategies and NMR methodologies to probe the architecture of model vaccine formulations at the atomic scale, in particular by probing the conformations of antigens within complex molecular assemblies, ii) applying them to samples of new peptide-based vaccine candidates for neurodegenerative diseases.

Activities

Preparation of labelled samples
Development and implementation of solid-state NMR experiments (including DNP)
Interpretation and enhancement of results
Contributing to the common tasks of the laboratory
Supervision of PhD students and trainees

Skills

Expertise in solid-state NMR spectroscopy
Expertise in biomolecular chemistry and structural biology

Work Context

The project will be developed at the Centre de RMN Très Haut Champ de Lyon, a research unit affiliated to the Ecole Normale Supérieure de Lyon, the CNRS (Centre National de la Recherche Scientifique) and the Université Lyon-1. Working environment : the CRMN hosts 7 NMR spectrometers of frequency 400, 500, 600, 700, 800 (x 2) and 1000 MHz, 2 gyrotrons for DNP MAS and 2 polarizers for dissolution DNP. Notably, the 1 GHz NMR spectrometer with 4 working MAS probes is fully operational for high-resolution solid-state NMR in chemistry and materials science. In particular, the spectrometer is equipped with a 0.7 mm MAS probe spinning at 110 kHz. The 800 MHz wide-bore spectrometer is equipped with a DNP instrumentation operating at 527 GHz (MW frequency) and three MAS probes of diameter 3.2, 1.3 and 0.7 mm and offers unique DNP capabilities. CRMN has a long-standing expertise for the structural and dynamic characterization of complex chemical and biological solids, including protein assemblies, fibrils and membrane proteins, heterogeneous catalysts, glasses, batteries and lighting materials, biomaterials, inorganic-organic photo- conducting devices, rocks and minerals. Its research groups are among the world leaders in fast MAS NMR, DNP enhanced solid-state NMR of surfaces, solid-state NMR of paramagnetic complexes. The site also has dedicated laboratories for sample preparation and handling.

Constraints and risks

None