Purification Process Design
Autor: Kristine Tjung • October 25, 2015 • Case Study • 1,565 Words (7 Pages) • 1,006 Views
Purification Process Design
Typical Purity Targets
- Primary Product Species > 95 %
- Product Related Species (Biologically Active) < 5 % total
- Oxidation (MetSO)
- Deamidation
- Minor N- or C-Terminal Truncation
- Reversible Aggregation
- Covalent Aggregate < 0.1%
- Unrelated Species < 0.1% each
- Nucleic Acid < 100 pg/dose
- Endotoxin < 1 EU/mg
- Adventitious Microbe none
- Adventitious Virus none
Chromatography Modes
Resin | Capacity (mg/ml) | Cost ($/L) | Cleaning Regimen | Lifespan (cycles) | Load Condition | Elute Condition |
Cation Exchange | 20-100 | Capture 500 Polish 3000 | 1) 1N NaAcetate 2) 1N NaOH | 100-1000 | Low-physiologic Salt, pH at least 1 unit below isoelectric point | High Salt or increased pH |
Anion Exchange | 20-100 | Capture 500 Polish 3000 | 1) 1N NaAcetate 2) 1N NaOH | 100-1000 | Low-physiologic Salt, pH at least 1 unit above isoelectric point | High Salt or decreased pH |
Hydrophobic Interaction | 2-10 | 1000-2500 | 1) Urea 2) 1N NaOH | 50 | High Precipitating Salt [1M (NH4)2SO4] | Low Salt, or Miscible Organic (ethylene glycol) |
Reversed Phase | 5-20 | 2000-5000 | High Organic (0.1% TFA, 100% Acetonitrile) | 50 | Low Organic (0.1% TFA, 10% Acetonitrile) | Intermediate Organic (0.1% TFA, 50% Acetonitrile) |
Metal Affinity | 5-50 | 1000-2500 | 1) EDTA 2) 1N NaOH | 100 | Neutral pH, 1M NaCl | Ammonium Chloride or Glycine |
Protein-A Affinity | 20-30 | 10000 | High Salt or GuHCl | 100-300 | Neutral pH, 0.15M NaCl | 1M Acetic Acid pH 3 |
Gel Filtration | 400-1000 | 0.5N NaOH | 50 | Concentrated | ~2-fold Dilution |
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