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

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

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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

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SLIDE 1

Processing and analysis of SEC-SAXS data

Alejandro Panjkovich

EMBL Hamburg

11.12.2017

  • A. Panjkovich (EMBL)

SEC-SAXS 11.12.2017 1 / 7

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SLIDE 2

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

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SLIDE 3

CHROMIXS: CHROMatography Inline X-ray Scattering

Visualize SEC-SAXS run Manually or automatic region selection Compare additional parameters and data Export data and figures Complete automatic processing

average intensity

8,500 9,000 9,500 10,000 10,500 11,000 11,500

radius of gyration

1 2 3 4 5 6

frame nr.

400 500 600 700 800 900 BSA SEC-SAXS monomer peak buffer Rg values

CHROMIXS

dimer peak

  • A. Panjkovich (EMBL)

SEC-SAXS 11.12.2017 3 / 7

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SLIDE 4

Experimental issues

  • A. Panjkovich (EMBL)

SEC-SAXS 11.12.2017 4 / 7

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SLIDE 5

Experimental issues

  • A. Panjkovich (EMBL)

SEC-SAXS 11.12.2017 5 / 7

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SLIDE 6

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

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SLIDE 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