Journal article

An optimized SEC-SAXS system enabling high X-ray dose for rapid SAXS assessment with correlated UV measurements for biomolecular structure analysis

Timothy M Ryan, Jill Trewhella, James M Murphy, Jeremy R Keown, Lachlan Casey, F Grant Pearce, David C Goldstone, Kelan Chen, Zhenyao Luo, Bostjan Kobe, Christopher A McDevitt, Serena A Watkin, Adrian M Hawley, Stephen T Mudie, Vesna Samardzic Boban, Nigel Kirby

JOURNAL OF APPLIED CRYSTALLOGRAPHY | INT UNION CRYSTALLOGRAPHY | Published : 2018

Abstract

A new optimized size exclusion chromatography small-angle X-ray scattering (SEC-SAXS) system for biomolecular SAXS at the Australian Synchrotron SAXS/WAXS beamline has been developed. The compact configuration reduces sample dilution to maximize sensitivity. Coflow sample presentation allows an 11-fold increase in flux on sample without capillary fouling, improving throughput and data quality, which are now primarily limited by the full flux available on the beamline. Multi-wavelength fibre optic UV analysis in close proximity to the X-ray beam allows for accurate concentration determination for samples with known UV extinction coefficients and thus estimation of the molecular weight of the ..

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Grants

Awarded by NHMRC


Awarded by National Health and Medical Research Council of Australia (NHMRC) RD Wright Fellowship


Awarded by NHMRC IRIISS


Awarded by NHMRC Programme


Funding Acknowledgements

DCG and JRK obtain support from the RSNZ Marsden fund. CAM was supported by NHMRC project grant 1080784. JMM was supported by a National Health and Medical Research Council of Australia (NHMRC) RD Wright Fellowship (APP1105754); KC was supported by a Cancer Council Victoria Postdoctoral Fellowship. JMM and KC's studies were enabled by NHMRC project grants 1045936 and 1098290, with additional support from NHMRC IRIISS 9000220 and Victoria Government Operational Infrastructure Support. BK is an NHMRC Principal Research Fellow (APP1110971) and the work was supported by NHMRC Programme APP1071659.