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Protein Expression and Purification Core Facility

PEPCF expresses proteins in bacteria, insect and mammalian cells and uses a variety of chromatographic and biophysical techniques for protein purification and characterization.

NMR support

NMR support provides assistance with nuclear magnetic resonance (NMR) spectroscopy across EMBL. The service supports researchers in the planning and execution of NMR experiments aimed at assessing biomolecular sample quality, probing molecular interactions, and characterising proteins, nucleic acids, small molecules, and their complexes.

NMR is a versatile technique for studying molecular structure, dynamics, and interactions in both liquid and solid environments. Measurements are coordinated through NMR support. For project discussions, experimental planning, and measurement coordination, please contact Alexander Marchanka (email-link here).

Available instrumentation

NMR experiments are carried out on a Bruker Avance III 700 MHz high-field spectrometer equipped with both solution-state and solid-state probes.

  • Solution-state NMR: room-temperature TXI triple-resonance probe (¹H/¹³C/¹⁵N)
  • Solid-state NMR: 3.2 mm MAS probe with triple-resonance capability (¹H/¹³C/¹⁵N)

Solution-state NMR applications

Solution-state NMR can provide valuable information on biomolecular and chemical samples in liquid phase. Depending on the scientific question and sample complexity, support can be provided at several levels:

  • Level 1 – 1D ¹H spectroscopy
    Rapid assessment of sample quality, purity, folding, and stability, with the possibility to obtain first insights into molecular interactions
  • Level 2 – 2D heteronuclear correlation experiments
    2D ¹H–¹⁵N HSQC and ¹H–¹³C HSQC experiments provide sensitive fingerprints of proteins, nucleic acids, and complexes, and are widely used for interaction analysis,  titration studies, and condition screening.
  • Level 3 – Advanced multidimensional NMR approaches
    For more complex projects (e.g. resonance assignments or higher-dimensional experiments), additional support and access pathways can be explored in collaboration with external partner infrastructures and NMR facilities, for example through Instruct-ERIC.

Further experiments can be discussed depending on project goals and feasibility.

Solid-state MAS NMR applications

Solid-state NMR is suitable for samples that are not accessible by solution methods, including insoluble or heterogeneous biomolecular systems. Possible applications include:

  • Spectral characterisation of biomolecular assemblies, complexes and aggregates
  • Advanced MAS-based ¹³C/¹⁵N solid-state NMR experiments for structural studies
  • 2D heteronuclear and homonuclear correlation measurements, depending on sample and feasibility

Solid-state measurements require early consultation, as sample preparation and optimisation are essential.

Sample requirements and preparation

Successful NMR experiments depend strongly on sample quality and preparation. Key aspects include:

  • Concentration, stability, and homogeneity of the sample
  • Buffer composition and suitability for NMR measurements
  • Isotopic labelling (¹³C/¹⁵N) for many biomolecular applications

Guidance on sample preparation and feasibility is provided as part of the support.

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