We provide a wide range of platforms at Diamond, from protein purification and characterisation (membrane proteins) to crystallisation screening and optimisation, data collection, and full structural analysis (available for both soluble and membrane proteins). You can choose to take up our crystallisation services at any stage of this process.
The success of membrane protein structural analysis largely depends on the quality of protein samples. We offer a comprehensive portfolio of biophysical methods to ensure high-quality control of protein samples. These methods help quickly identify the most suitable protein constructs and purification conditions, followed by thorough protein characterisation. Proprietary clients can enter the pipeline at any stage, provided the protein sample passes the quality control step, confirming its suitability for subsequent steps.
Protein samples undergo high-throughput extraction and purification screens. Optimal conditions are selected based on SDS-PAGE and Fluorescence-detection Size Exclusion Chromatography (FSEC) results.
Membrane protein purification and formulation allow scientists to screen a wide range of conditions, including detergents, amphipols, membrane scaffold proteins (MSPs), SMALPS, lipids, pH, and salts. A typical purification process involves Immobilised Metal Affinity Chromatography (IMAC) using His-tag, reverse IMAC after construct cleavage (TEV or 3C protease), and Size Exclusion Chromatography (SEC).
Protein quality is confirmed using SEC-MALS and a thermostability assay, provided the protein is sufficiently pure (>90%). If a simple assay is available, the target protein function can be tested at this stage. For example, if the target protein substrate is known, Nano DSF can be used to check for a protein-substrate interaction.
The Crystallisation Facility (CF) at Diamond offers comprehensive services for crystallisation and crystal harvesting for X-ray diffraction data collection. Co-located with the Diamond synchrotron, CF services are integral to the full-service structural biology package available to industrial customers, including X-ray crystallography and structural analysis of protein targets.
High-throughput robotics at the CF facilitate the crystallisation of both soluble proteins and membrane proteins in Lipidic Cubic Phase (LCP). Crystallisation screening is available using common commercial coarse screens and custom screens prepared with the Formulator liquid handler. Robotics such as Mosquito (SPT Labtech) and Gryphon (Art Robbins) are used for preparing sitting drop and LCP crystallisation plates.
The Formulatrix Rock Imager 1000 imaging systems offer a range of crystallisation temperatures and feature cross-polarisers, UV, and multi-fluorescence imaging options. These images can be accessed remotely.
Crystallisation of soluble proteins
Purified and characterised proteins can be used for initial crystallisation screening using commercially available screens. Vapour diffusion crystallisation can be conducted at various temperatures, including: 4oC, 12oC, 16oC, 19oC and 20oC.
Examples of the commercial screens offered at the platform include:
