In-Lab SEC-SAXS for Structual Investigation of Protein Samples

  • SAXS can be used to investigate biological macromolecules especially in monodisperse solutions.

    Xenocs BioXolver equipped with MetalJet

  • However liquid bio-solutions involve structurally different molecules and complexes in dynamic equilibria.
  • SEC – Size Exclusion Chromatography has evolved as a technique to mitigate this by separating the molecules in time.
  • However this limits the measurements time window which put huge
    requirements on the X-ray source.
  • With MetalJet source in the Xenocs BioXolver system it is possible to
    perform protein in-lab SEC-SAXS even for very diluted solutions containing
    less than 1 mg of sample.

   

Application note kindly provided by Application Scientist at Xenocs SAS using a BioXolver equipped with Metaljet X-ray source.
You find the Appl. note in full here.
Bucciarelli, S. et al. Size-exclusion chromatography small-angle X-ray scattering (SEC-SAXS) of water soluble proteins on a laboratory instrument, J. Appl. Crystallogr. 2018

Fibres

Application scientists at Bruker AXS have used a NANOSTAR instrument equipped with a MetalJet (200W at 70kV) to record a series of SAXS scattering plots of a very thin fibre from a rat tail tendon. Providing intensity gains of more than 50x compared to state-of-the-art sealed microfocus tubes.

 

 

Biological (BioSAXS)

Researchers at Nanyang Technical University, A*STAR, University of Louisville, Rosalind Franklin University of Medicine and Science and Kyung Hee University have used a MetalJet equipped SAXS instrument to study the BclxL protein, a key regulator in mitochondrial calcium ion transport. The protein was treated with a mild detergent and studied during the formation of a dimer resulting from three-dimensional domain swapping (3DDS) of helices α6-α8 between two monomers.

Sci. Rep. 5, 10609 (2015), S. Rajan, M. Choi, Q. T. Nguyen, H. Ye, W. Liu, H. T. Toh, C. B. Kang, N. Kamariah, C. Li, H. Huang, C. White, K. Baek, G. Grüber, H. S. Yoon.

 

 

Metals and colloids

Researchers at the Slovak Academy of Science and STU Centre for Nano-diagnostics performed in-situ tests on a strain gauge, based on a monolayer of colloidal gold nanoparticles deposited on a flexible Mylar foil. The tests were monitored by SAXS where the high brightness of the MetalJet allowed a very fast data collection, with 10 seconds temporal resolution.

Sensors and Actuators A: Physical, 2016, 247, 87-95, K. Vegso, M. Jergel, P. Siffalovic, M. Kotlar, Y. Halahovetsa, M. Hodasa, M. Pellettaa, E. Majkovaa

 

 

Polymers

The investigation of polymers is one of the main areas of focus of the Soft Matter Analytical Laboratory (SMALL) at the University of Sheffield. Following a £2 million infrastructure investment in 2016 and the purchase of a new SAXS system with a gallium MetalJet X-ray source, their first new resulting publication was a co-polymerisation study of Styrene with N-Phenylmaleimide.

Macromolecules, 2016, 49 (18), 6731–6742, P. Yang, O. O. Mykhaylyk, E. R. Jones, S. P. Armes.

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