Protein SAXS (BioSAXS)

Why choose BioSAXS at Diamond

Small angle X-ray scattering (SAXS) from biological macromolecules such as proteins (also called bioSAXS) gives information on the size and shape of macromolecules in solution. It is increasingly used by the pharmaceutical industry to provide details of the radius of gyration and molecular weight of a protein complex (e.g. multi sub-unit complexes), significant conformational changes upon ligand binding, screening of structural changes with formulation conditions (e.g. pH, ionic strength, additives, temperature etc) and many more applications.

With high throughput and fast and reliable measurements, bioSAXS at Diamond enables low volume samples to be studied at close to native state conditions. It is a very effective tool in situations where crystals are unattainable or where samples may be challenging eg. flexible proteins, or multi-sub unit complexes (e.g antibody binding studies). The highest quality data can be achieved using online size exclusion chromatography (SEC-SAXS), ideal for advanced modelling. Online MALS is also available, allowing validation of laboratory based studies.

Often used in conjunction with structural information from crystallography or electron microscopy bioSAXS may form part of a wider set of research, made simple by the co-location of Diamond’s broad portfolio of synchrotron and cryoEM techniques, all housed on one central site.

Our expert bioSAXS team can support and advise you in how to get the best from your research using this technique and are on hand to support you throughout your experiment.

Benefits

Of BioSAXS research technique

Allows studies in near native state conditions
Accommodates challenging systems (eg. flexible proteins)
Allows small volumes (~35 µL) to be measured
Provides unique information about the shape and size of macromolecules
Gives an average structure from a bulk sample i.e. complementary to Electron Microscopy studies

Of synchrotron BioSAXS

Batch mode data collection gives very high throughput
Fast measurement times
Low solution volumes
Can achieve a wider range of length scales compared to lab sources
Can be performed in conjunction with other techniques (online MALS)
Provides insight under different formulation conditions (eg. temperature, pH)
SEC-SAXS mode data collection provides high purity samples for optimum data quality

Of BioSAXS at Diamond:

Dedicated state-of-the-art beamline
Streamlined automatic data processing
Dedicated analysis software freely available for industrial use
Unique modelling & advanced analysis support
Industry specific expertise and support
Colocation with other research techniques
Full automated data collection modes (BioSAXS robot and SEC-SAXS) that can be used in combination

Applications

Probe microstructures at mesoscopic scale from polypeptides to large complexes
Determine oligomerisation state of a protein or protein complex (eg. for targeted antibodies)
Characterise multi-domain proteins using data from subcomponents of a modular protein or complex
Probe conformational changes associated with ligand binding.
Investigate flexible proteins & samples non amenable to crystallisation
Determine suitable conditions for crystallisation
Validate crystal structures
Facilitate large molecule drug discovery
Determine changes due to external environments (eg. pH and temperature, and the effects of excipients, coatings and medical devices)

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Protein SAXS (BioSAXS)

Why choose BioSAXS at Diamond

Benefits

Applications