Ligand Binding Studies
NMR binding studies, ranging from a single experiment to a consensus-based approach combining various experiments. Compounds are ranked by scores to guide medicinal chemistry efforts.
⦾ 1H Ligand-Detection
Provides a thorough evaluation of target-compound binding, sample integrity, compound concentration and binding epitope(s).
⦾ 19F Ligand-Detection
The highly sensitive fluorine atom allows for fast, background-free detection of a wide range of binding events.
⦾ Protein-Detected NMR
Higher-affinity compounds can sometimes be missed by ligand-observed techniques; Monitoring protein fingerprints helps to bridge the gap between ligand-detected NMR and other biophysical techniques, while keeping track of protein integrity.
Kd Determination
Binding events are detected by the perturbation of the target protein residues. Residues are tracked via NMR during titration experiments with the ligand of interest, where Kd’s in the mM to very low μM range can be determined, even without knowing the identity of the perturbed residues. Orthogonal methods are performed to corroborate Kd by NMR, including Surface Plasmon Resonance, Grating Coupled Interferometry, or Isothermal Titration Calorimetry.
⦾ Kd & Stoichiometry
Extraction of binding affinity constants from either protein-detected or ligand-detected NMR experiments.
Compound Behaviour (Free State Properties)
Understanding behaviour of compounds in relevant conditions to guide experimental testing, controlling for a major source of artifacts.
⦾ Aggregation
Relaxation, dilution, and detergent effect assays are performed to get the most accurate picture of the behaviour of relevant compounds in solution.
⦾ Accurate Concentration Determination
The actual compound concentration of each sample is systematically determined, using data obtained during any given NMR experiment. It can also be independently measured as part of a solution behaviour routine.
Target Enablement
Characterization of the target protein folding, stability, promiscuity, and optimal concentration for the screening process by 1D NMR assays. It is crucial to ensure the target is well-behaved and optimal screening conditions are defined.
Fragment Library Screening
1H and 19F screen of low molecular weight fragments by NMR. Hits are then ranked according to consensus binding between different NMR experiments. Clients can avail of NMX’s curated and ready-to-go libraries, or can opt to bring their own!
⦾ Curated Fragment Librairies
Our 1H and 19F fragment libraries have gone through rigorous cheminformatics filtering and exhaustive NMR curation in buffer to retain only well-behaved and drug-like compounds.
⦾ Optimized Pooling
Our curated fragment libraries are screened as compatible mixtures of fragments, in order to increase throughput, allowing for a streamlined evaluation of the binding of thousands of drug-like fragments to your target.
Binding Site Identification and 3D Structures
Mapping the binding site(s) of compounds of interest on a target, using direct or indirect NMR experiments, along with information from computational modeling and/or X-ray crystallography.
⦾ Chemical Shift Mapping
Direct identification by NMR of perturbed target residues upon addition of a compound of interest.
⦾ Competition Assays with Tool Compounds
Inference of the binding site of a molecule using competition assays with tool compounds of known binding modes.
