OCT 14, 2014 01:00 AM PDT

Aggregation Analysis and Beyond: Analytical Ultracentrifugation in the Biopharmaceutical Industry

Speakers
  • Head of the AUC facility of Sandoz/Novartis
    Biography
      Alexander Bepperling graduated with a diploma in biochemistry in the lab of Prof. Rainer Rudolph (University Halle Institute for industrial Biotechnologie) and obtained his PhD in biotechnology in the group of Prof. Johannes Buchner (Technical University Munich, CIPSM: Center of Integrated Protein Research Munich). He worked for almost 10 years in the field of biophysics with special focus on analytical ultracentrifugation. He joined Sandoz in 2011 as a scientist and is currently heading the AUC facility of Sandoz/Novartis but employing also various spectroscopic technologies (CD, FTIR, Fluorescence spectroscopy) as well as surface plasmon resonance spectroscopy. 
       

    Abstract:

     

    AUC sedimentation velocity (SV) has a longstanding reputation as being the gold standard for the matrix free quantification of aggregates in biopharmaceuticals. The continuous c(S) distribution analysis as implemented in the software SEDFIT is the most commonly used data evaluation method in the biopharma environment due to regulatory requirements and the reliable reporting of very minor species by the algorithm. I will show how to set up such an analysis in order to achieve enhanced reproducibility during a study of many batches of one protein. Attention will be also paid to the recently developed calibration procedures regarding the temperature, scan time and the radial calibration of the centrifuge.
    In the second part examples are shown apart from the generic method of quantitating aggregates in dilute solutions under standardized conditions. These examples will include the determination of the extinction coefficient and concentration of protein formulations, a task far from being trivial in the biopharmaceutical field and the determination of the second virial coefficient, which is known as a predictor for aggregation propensity. The presentation will conclude with some applications of sedimentation equilibrium (SE) for the screening of weak interactions and the determination of detergent binding to proteins.

    Learning Objectives:

    • Learn how to set up continuous c(S) distribution analysis as implemented in the software SEDFIT in order to achieve enhanced reproducibility during a study of many batches of one protein
    • See some applications of sedimentation equilibrium (SE) for the screening of weak interactions and the determination of detergent binding to proteins

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