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Biopharmaceuticals offer great new possibilities and potential in treating various life threatening diseases such as cancer and autoimmune diseases. However, their complexity, resulting from their heterogeneity, represents a great challenge to the analytical chemist, and a suite of techniques is required in the characterization and analysis. In conjunction with RP and HILIC, covered in Part I, ion exchange chromatography (IEX), and size exclusion chromatography (SEC) are the main stay of the biopharmaceutical analyst’s arsenal. These techniques provide much more information than merely charge variance and aggregation analysis. In this event we will discuss how, why and where IEX and SEC are typically employed in biopharmaceutical characterization and quantification. We will also highlight strategies for developing more robust IEX and SEC methods such as reducing SEC secondary interactions and contrast the use of pH and salt gradients in IEX. We will also explore the affect temperature and pressure have on SEC separations and the role of UHPLC. An introduction to hydrophobic interaction chromatography (HIC) will also be given.

 

Topics include:

  • Why is it necessary to use size exclusion (SEC) and ion exchange (IEX) chromatography
  • What information can be gleaned from SEC and IEX separations?
  • How common are secondary interactions in SEC and how can they be minimized?
  • How do pH gradients compare with traditional salt gradients in IEX?
  • What impact do temperature and pressure have on aggregation?
  • Does UHPLC have a key role in SEC separations?