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Hydrophobic Interaction Chromatography Column

Hydrophobic Interaction Chromatography Column. Stephanie Budiwarman Steven O’Neill Leanna Yue. Remove water, conc cell suspension, lyse and homogenize . GM E .coli is multiplied. Trp - Proinsulin. Proinsulin –SSO3. Human insulin. Proinsulin. Background Theory.

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Hydrophobic Interaction Chromatography Column

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  1. Hydrophobic Interaction Chromatography Column Stephanie Budiwarman Steven O’Neill Leanna Yue

  2. Remove water, conc cell suspension, lyse and homogenize GM E .coli is multiplied Trp - Proinsulin Proinsulin –SSO3 Human insulin Proinsulin

  3. Background Theory

  4. Operational Parameters • Choice of medium • Ligand and bead size • Choice and type of eluent • Step vs. Gradient • Temperature, Pressure • Column size • Flow rate

  5. Final Design • Medium • Octylsepharose (90μm beads) • Eluent • Step elution; NaCl • Temperature and Pressure • 25oC, 1atm • Size • H= 0.25m, D= 2.093m • Flow Rate • 175 – 200 cm/h Surface of agarose bead OctylSepharose

  6. Suppliers and Costing • GE Healthcare • Medium: $800 for 200mL • Shodex • Tosho Bioscience LLC • Column: $1200 lab equipment

  7. Alternative Methods • Affinity Chromatography (AF) • Specific ligand • Ion Exchange (IEX) • Ionic charge • Reverse Phase Chromatography (RPC) • Hydrophobicity • Solvent denatures proteins • Gel Filtration (GF) • Molecule size • Low resolution

  8. Questions?

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