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

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Processing and analysis of SEC-SAXS data Alejandro Sasha Panjkovich Celgene Corporation 2019.10.19 A. Panjkovich (Celgene) SEC-SAXS 2019.10.19 1 / 7 Outline Visualize elution profile in terms of scattering Common experimental issues

Processing and analysis of SEC-SAXS data Alejandro ‘Sasha’ Panjkovich Celgene Corporation 2019.10.19 A. Panjkovich (Celgene) SEC-SAXS 2019.10.19 1 / 7
Outline Visualize elution profile in terms of scattering Common experimental issues Identify sample and buffer region Evaluate sample behaviour (Rg and MW) Align with additional data A. Panjkovich (Celgene) SEC-SAXS 2019.10.19 2 / 7
CHROMIXS: CHROMatography Inline X-ray Scattering CHROMIXS BSA SEC-SAXS Visualize SEC-SAXS run 6 11,500 monomer peak dimer peak Manual 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 (Celgene) SEC-SAXS 2019.10.19 3 / 7
Experimental issues A. Panjkovich (Celgene) SEC-SAXS 2019.10.19 4 / 7
Experimental issues A. Panjkovich (Celgene) SEC-SAXS 2019.10.19 5 / 7
Reference and other programs for further analysis CHROMIXS (part of ATSAS package) Panjkovich and Svergun (2018) CHROMIXS: automatic and interactive analysis of chromatography-coupled small angle X-ray scattering data. Bioinformatics 34:1944-46 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, 1545–1553. Scatter http://www.esrf.eu/UsersAndScience/Experiments/CRG/BM26/SaxsWaxs/DataAnalysis/Scatter A. Panjkovich (Celgene) SEC-SAXS 2019.10.19 6 / 7
Summary Visualize elution profile in terms of scattering Common experimental issues Identify sample and buffer region Evaluate sample behaviour (Rg and MW) Align with additional data A. Panjkovich (Celgene) SEC-SAXS 2019.10.19 7 / 7
EMBO Global Exchange Lecture Course Structural and biophysical methods for biological macromolecules in solution 14 – 20 October 2019 | Santiago, Chile

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