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Industrial Liaison Group:
Tel: +44 (0) 1235 778797
E-mail: industry@diamond.ac.uk
Porton Biopharma Ltd, first industrial users on B21, have today published their work in Pharmaceutical Research.
Dr Dave Gervais and colleagues from Porton Biopharma Ltd worked in collaboration with Diamond's Rob Rambo, Principal Beamline Scientist for B21 and Claire Pizzey, Deputy Head of Industrial Liaison, and scientists from the Institute of Structural and Molecular Biology, UCL and the Department of Chemical and Biological Engineering, University of Sheffield on the structural characterisation of biopharmaceuticals.
The biopharmaceutical of interest in this work is an enzyme called erwinia chrysanthemi L-asparaginase (ErA), used in the treatment of acute lymphoblastic leukaemia. The paper has shown that biopharmaceutical products such as ErA should be carefully monitored for common modifications, such as deamidation, using ion-exchange chromatography.
The paper highlights the analysis of clinical grade ErA using native ion exchange chromatography (IEX), showing that a number of enzymatically active, acidic variants were present that previously had been poorly characterised. In this study, the variants were isolated and fully characterised using a range of biophysical techniques. Structural analysis of the variants was performed using small angle X-ray scattering (SAXS), circular dichroism and ion-mobility mass spectrometry (IM-MS). The SAXS experiments performed at Diamond revealed a different conformational state in the most acidic variant (from IEX), also suggested by the IM-MS data.
The SAXS experiments also suggested that minor conformational differences between the main peak from IEX and the major acidic variant existed, based on comparison with crystal structures.
Full details of the experiments can be found in the paper and for further information about biological SAXS experiments at Diamond, please give us a call or send us an e-mail, we'd love to hear from you.
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