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BIT 464/564: Protein Purification


  • Introduce students to basic skills and theory needed purify recombinant proteins from microbial expression hosts using FPLC
  • Use enzymes from hyperthermophiles to simplify assays and minimize stability problems


  1. Introduction to Liquid Chromatography
  2. Protein Purification Basics
  3. Liquid Chromatography – Ion Exchange
  4. Liquid Chromatography – Hydrophobic Interaction
  5. Liquid Chromatography – Gel Filtration/Affinity
  6. Protein Folding and Re-folding
  7. Downstream Processing


  1. FPLC Operation – Purification of Protein Standards 1 week  
  2. AEC purification of Thermotoga maritima -gal from Heat-Treated E. coli 1 week  
  3. Purification of Thermotoga maritima -gal from E. coli by AEC, HIC, HAP 3 weeks
  4. Purification of Thermotoga maritima -gal from E. coli by His-tag/IMEC 1 week  
  5. Finish assays and run gels 1 week  

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