Structural Biology Services | Sapient Discovery

Structural Biology Services

Sapient Discovery offers structural biology contract and consultant services as a cost-effective and powerful approach to help validate and guide medicinal efforts in the drug design and optimization process. SD's structural biology facility provides protein production and X-ray crystallography services with end result of providing at atomic level a 3D snapshot of the small molecule with the target protein. The structural details of interactions and conformations of protein target and co-crystal structures of target with customer's small molecule provides structural insights for exploitation of the target for the design of more select and potent inhibitors that custom-fit into the protein target of interest.

Protein Engineering

Construct Design: At Sapient Discovery we first use bioinformatic tools to design protein constructs that are suited for the structural studies. Using structural and function information constructs are designed and engineered. N- and or C-terminal peptide/protein tags are engineered to help in the expression and protein purification stages.

Mutations: Amino acid mutations are engineered into the target protein to improve solubility and crystallization or help in further elucidating the structure activity relationship and mode of interaction of the inhibitor.

Selenomethionine Incorporation: Sapient Discovery can generate selenomethionyl derivatives of the target protein in instances where X-ray structure elucidation requires additional electron density improvement or phase determination via multi-wavelength anomalous dispersion (MAD).

Cloning & Expression

A handful of designed protein constructs encompassing the structural domain of interest are cloned and expressed in several systems. Multiple expression systems and conditions are performed in small-scale pilot tests to determine the optimal expression of soluble protein for large-scale protein production.

Protein Purification

Proteins are purified to crystallography grade using one or more purification steps.

Protein Production Services

Sapient Discovery's protein production capabilities could be accessed for producing novel proteins as a fee-for-service. Proteins are routinely expressed and purified in our laboratory in milligram quantities for crystallography studies.

Sapient Discovery's proprietary bioinformatics approach helps define a functional domain best suited for expression & crystallization.

Amino acid mutations can be engineered into the target protein to improve solubility and crystallization or help in further elucidating the structure activity relationship.

Select constructs are cloned and expressed. Multiple expression systems and conditions are performed in small-scale pilot tests to determine the optimal expression.

Protein is expressed in large scale under optimal expression conditions.

Target protein is purified in milligram quantity to crystal grade for structure determination studies.

X-ray Crystallography Services

Sapient Discovery offers X-ray crystallography services to accelerate customer's pre-clinical drug discovery programs providing a powerful tool at both the hit-to-lead and lead optimization stages. We are now seeking companies who might have a need for X-ray crystallographic services to work with us on a fee-for-service mode, to bring protein crystal structures to their drug-discovery/vaccine research/protein engineering efforts.

Sapient Discovery uses its proprietary crystallization screening conditions at both the pilot and scale-up crystallization stages. The small molecule candidate is either co- crystallized with the protein or is soaked into the final protein crystal in a temperature-controlled environment.

Pure target protein is generated in milligram quantity for crystallization.

Pilot crystals are obtained using SD's proprietary crystallization screening methods.

Crystallization conditions are optimized to generate data quality crystals. Crystals can either be grown in the presence or absence of the small molecule.

Crystals are grown and stored in a temperature controlled environment.

Crystals are flash cooled for X-ray data collection.

X-ray Diffraction data set is collected from a cryo-frozen protein crystal.

X-ray data is processed and electron density maps calculated for the protein and the small molecule.

Sapient Discovery has all the state of the art computational infrastructure, crystallography software and high resolution interactive graphic systems for data processing, reduction, structure determination, refinement, fitting and viewing of electron density maps and structures.

The final X-ray structure provides at atomic level a 3D snapshot of the small molecule bound to its the target protein. Co-crystal structures can aid in the design of more select & potent inhibitors.