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Ion Exchange Chromatography

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Learn the key principles of ion exchange chromatography, and the relationship between pH, protein and isoelectric points (pl). Examine an ion exchange column to predict which proteins will bind or elute under differing pH conditions.

Learning Outcomes

  • Consider the relationship between pH, protein isoelectric points (pI) and protein charge
  • Explore how differences in pI of proteins can be used to aid protein purification
  • Predict protein binding or elution to anion and cation exchange columns under different pH conditions
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Concepts Principles and Theories

  • What is the relationship between pH and overall charge on a protein
  • What is meant by the isoelectric point
  • How differences in the isoelectric point of proteins can be used to aid protein purification
  • How to predict whether a protein will bind to the column equilibrated at a particular pH when given its isoelectric point
  • How changing the buffer added to the column affects the pH environment and elution of proteins

Designing and Planning Experiments

  • What is the difference between a cation and anion exchange column

Interpreting and Analysing Data

  • How to identify the order proteins will reach their isoelectric point when given a graph of charge vs pH

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