NMR-Based Fragment-screening in GUF
Service description:
BMRZ is a major hub for RNA/protein structural biology, offering 20+ years of expertise in fragment-based NMR screening. As a core service, BMRZ provides ligand (1H)-observed fragment screening to identify binding partners for diverse biomolecules. Within the Fragment-Screen project (WP2/6), GUF developed an optimized protocol using the DSi-poised library in mixtures of four on a 600 MHz spectrometer with a 1.7 mm probe, now available for pilot studies. Since 2020, BMRZ has screened 45+ pandemic-related targets (RNA and proteins), refining high-throughput workflows and semi-automated analysis. Two dedicated spectrometers support specialised 19F and low-gamma detection, providing users access to libraries of 600-1000 molecules.
User guideline:
This pilot study is designed for soluble protein or RNA targets, which will be screened against the DSi-poised library formatted into 48 mixtures (four fragments per mixture). To ensure high-quality, error-free analysis and minimise false positives, the following technical criteria apply:
- Sample Requirements: The user must provide 10 ml of the target (10 µM) in an optimised buffer; preferably a phosphate buffer at pH 7.4. Crucially, 10 ml of the final exchange buffer must also be provided for blank NMR measurements.
- Technical Setup: Screening will be conducted as a ligand-observed service on a 600 MHz spectrometer equipped with a Bruker SampleJet and a high-sensitivity 1.7 mm probe.
- Workflow & Timeline: The total duration for sample preparation and NMR measurements (including 1D, STD, and WaterLOGSY experiments) is 4–5 days. Facility experts will provide the user with comprehensive guidelines for subsequent data analysis.
- Consultation: Users are strongly encouraged to consult the facility manager before finalising buffer conditions. Pre-screening coordination ensures optimal conditions, avoids tedious data deconvolution, and significantly improves hit-validation efficiency.
- Expected timeline for the access visit: May-June 2026
