You can use the CiPP (capture, intermediate purification, polishing) purification strategy to structure your purification protocol:

   • The initial capture stage isolates, concentrates, and stabilizes your protein.
   • Intermediate purification removes bulk contaminants.
   • The final polishing step removes the most difficult impurities, such as aggregates of the target protein.

In a research setting, purity requirements are often less important than yield and you may therefore only need a chromatography capture step to achieve your goal.

A general rule of thumb is that the higher protein purity you need, the more purification steps you will have to deploy in your workflow. However, additional chromatography steps will increase purity but decrease yield of active protein.