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BIT 464/564: Protein Purification

Overview

  • ntroduce 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

Lectures:

  • Introduction to Liquid Chromatography
  • Protein Purification Basics
  • Liquid Chromatography – Ion Exchange
  • Liquid Chromatography – Hydrophobic Interaction
  • Liquid Chromatography – Gel Filtration/Affinity
  • Protein Folding and Re-folding
  • Downstream Processing

Labs:

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