Processing and analysis of SEC-SAXS data Alejandro Panjkovich EMBL - PowerPoint PPT Presentation

Processing and analysis of SEC-SAXS data Alejandro Panjkovich EMBL Hamburg 11.12.2017 A. Panjkovich (EMBL) SEC-SAXS 11.12.2017 1 / 7

Outline Visualize elution profile in term of scattering Common issues Identify sample and buffer region Evaluate sample behaviour (Rg and MW) Align with additional data A. Panjkovich (EMBL) SEC-SAXS 11.12.2017 2 / 7

CHROMIXS: CHROMatography Inline X-ray Scattering CHROMIXS BSA SEC-SAXS Visualize SEC-SAXS run 6 11,500 monomer peak dimer peak Manually or automatic 11,000 5 buffer Rg values region selection 10,500 average intensity 4 radius of gyration Compare additional 10,000 3 parameters and data 9,500 2 Export data and figures 9,000 1 Complete automatic 8,500 processing 400 500 600 700 800 900 frame nr. A. Panjkovich (EMBL) SEC-SAXS 11.12.2017 3 / 7

Experimental issues A. Panjkovich (EMBL) SEC-SAXS 11.12.2017 4 / 7

Experimental issues A. Panjkovich (EMBL) SEC-SAXS 11.12.2017 5 / 7

Reference and other programs for further analysis CHROMIXS (part of ATSAS package) Panjkovich and Svergun (2017) CHROMIXS: automatic and interactive analysis of chromatography-coupled small angle X-ray scattering data. Submitted US SOMO Brookes et al. (2016) ”US-SOMO HPLC-SAXS Module: Dealing with Capillary Fouling, and Extraction of Pure Component Patterns from Poorly Resolved SEC-SAXS Data”, J. Appl. Cryst. 49. BioXTAS RAW Hopkins, J. B., Gillilan, R. E., and Skou, S. (2017). Bioxtas raw: improvements to a free open-source program for small-angle x-ray scattering data reduction and analysis. Journal of applied crystallography, 50, 15451553. Scatter http://www.esrf.eu/UsersAndScience/Experiments/CRG/BM26/SaxsWaxs/DataAnalysis/Scatter A. Panjkovich (EMBL) SEC-SAXS 11.12.2017 6 / 7

Summary Visualize elution profile in term of scattering Common issues Identify sample and buffer region Evaluate sample behaviour (Rg and MW) Combine with additional data A. Panjkovich (EMBL) SEC-SAXS 11.12.2017 7 / 7