Construct Design and Selection
The design of suitable constructs is key for a successful structure determination for your target protein. Based on target experience, bioinformatic analyses, and limited proteolytic digest analyses, we identify a set of different-length construct variants suitable for addressing the aims of your program. Our protein science labs perform expression and purification tests to identify the optimal conditions for target protein production. Biophysical methods (including TSA and nanoDSF) may be used to identify optimal constructs, buffer composition, or early hit list compounds that stabilize the target protein and facilitate crystallization.
Crystallization Screening
Our state-of-the-art crystallization platform, which enables fast and efficient screening to identify and optimize crystallization conditions, includes:
- crystallization screen preparation using Freedom EVO (Tecan) and FORMULATOR (FORMULATRIX) liquid-handling systems
- broad panel of proprietary and vendor screens, including protein-class-specific sparse-matrix crystallization screens (e.g., antibody screens)
- accurate drop setting using Mosquito nanoliter liquid-handling machines (including humidity control box)
- specialized membrane protein liquid cubic phase (LCP) crystallization platform, including an LCP mixing station and an LCP-enabled Mosquito nano-dispensing robot
- specialized fragment-screening platform, including an Echo Acoustic Liquid Handler for soaking experiments and a Crystal Shifter for crystal freezing
- automated crystallization imaging (visible and UV) at 4°C and 20°C using the FORMULATRIX ROCK MAKER imaging systems
Data Collection, Structure Determination, and Analysis
Regular bi-weekly access to third-generation high-brilliance synchrotron sources is complemented with our in-house automated crystal screening sample changer and two X-ray sources. Together, they support rapid and regular data collection. Our in-house X-ray platform includes:
- Rigaku MicroMax rotating anode generator (007HF) equipped with a PILATUS 200 detector and ACTOR robot for automated sample mounting
- Rigaku XtaLAB Synergy-S generator equipped with a HyPix-6000HE detector
Supported by our proprietary automated structure-solution pipeline, datasets are evaluated and structures are fully refined. Following a final internal quality control check, we deliver a structural biology data package containing a full set of industry-standard files.
Together with our DLS team, we complete the structure-determination activities by offering an in-depth binding mode analysis to support and accelerate structure-based compound optimization and to help enable full integration of structural learnings into your optimization programs.
References
Günther, J., et al., BAY-069, a Novel (Trifluoromethyl)pyrimidinedione-Based BCAT1/2 Inhibitor and Chemical Probe. J Med Chem (2022)
Orsi, DL., et al., Discovery and characterization of orally bioavailable 4-chloro-6-fluoroisophthalamides as covalent PPARG inverse-agonist. Bioorg Med Chem (2023)
Hillig, RC., et al., Discovery of potent SOS1 inhibitors that block RAS activation via disruption of the RAS-SOS1 interaction. PNAS (2019)
Hahn, MG., et al., Discovery of the Soluble Guanylate Cyclase Activator Runcaciguat (BAY 1101042). JMedChem (2021)
Roehrig et al., Design and Preclinical Characterization Program toward Asundexian (BAY 2433334), an Oral Factor XIa Inhibitor for the Prevention and Treatment of Thromboembolic Disorders. JMedChem, 2023, 66 (17)
