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BioPro IEX SmartSep Q/S are ion exchange media for high-throughput intermediate purification step and polishing step of biopharmaceuticals. BioPro IEX SmartSep Q/S are available as strong ion exchangers of hydrophilic porous polymer beads with low nonspecific adsorption and high binding capacity over a wide range of flow rates. BioPro IEX SmartSep Q/S show high resolution and recovery, even at high flow rates and high loading conditions.
| BioPro IEX SmartSep Q | BioPro IEX SmartSep S | |
|---|---|---|
| Matrix | Hydrophilic porous polymer | |
| Particle size (μm) | 10, 20, 30 | |
| Charged group | -R-N+（CH3）3 | -R-SO3- |
| Usable pH range | 2.0 - 12.0 | |
| Ion exchange capacity （meq/mL-resin） | >0.08 | |
| Dynamic binding capacity (DBC) (mg/mL-regin) | DBC >100 (BSA) | DBC >100 (lysozyme) |
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BioPro IEX SmartSep Q/S have higher DBC than conventional ion exchange media. In particular, for IgG, the DBC of BioPro IEX SmartSep Q/S is more than twice as high as that of the competitor.
This feature of BioPro IEX SmartSep Q/S makes purification productivity of IgG per unit time double or more.
Conditions of DBC measurement *
| Column | ： | 50 X 5.0 mmI.D. |
|---|---|---|
| Flow rate | ： | 400 cm/hr (1.32 mL/min) |
| DBC (mg/mL-resin, 10% breakthrough) | |||
|---|---|---|---|
| Insulin | Lysozyme | Human Polyclonal IgG | |
| BioPro IEX SmartSep S30 | 73 | 111 | 93 |
| Brand T (Porous S type, 30 µm) | 67 | 72 | 42 |
| Brand G (Porous S type, 30 µm) | 64 | 85 | 41 |
As the high DBC of BioPro IEX SmartSep Q/S is maintained at higher flow rates, it is suitable for high-speed purifications with 2–4 times the conventional flow rate. This feature offers a significant improvement of productivity.
| Equilibration buffer | ： | 20 mM citric acid-NaOH (pH 5.3) |
|---|---|---|
| Elution buffer | ： | Equilibration buffer cont. 0.5 M NaCl |
| Flow rate | ： | 200-800 cm/hr (0.66-2.62 mL/min) |
| Sample | ： | 1.5 mg/mL human polyclonal IgG in equilibration buffer |
BioPro IEX SmartSep Q/S show high resolution and recovery, even at high flow rates and high loading conditions. BioPro IEX SmartSep Q/S offer high efficiency for intermediate purification step and polishing step that require high resolution and recovery.
Separation at a high Flow Rate and High Loading Condition
| Column | : | 50 X 5.0 mmI.D. |
|---|---|---|
| Eluent | : | A) 20 mM NaH2PO4-Na2HPO4 (pH 6.8) B) 20 mM NaH2PO4-Na2HPO4 (pH 6.8) containing 0.5 M NaCl 0-100%B, 30 column volumes |
| Flow rate | : | 1600 cm/hr (5.23 mL/min) |
| Temperature | : | 25ºC |
| Detection | : | UV at 220 nm |
| Injection | : | 30 mL (45 mg Proteins) |
| Sample | : | 1. Ribonuclease A (0.5 mg/mL) 2. Cytochrome c (0.5 mg/mL) 3. Lysozyme (0.5 mg/mL) |
| Recovery (99% Purity) | ||||
|---|---|---|---|---|
| Ribonuclease A | Cytochrome c | Lysozyme | Total | |
| BioPro IEX SmartSep S30 | 90.94% | 80.33% | 99.18% | 90.64% |
| Brand T (Porous S type, 30 µm) | 80.64% | 59.60% | 98.28% | 80.08% |
| Brand G (Porous S type, 30 µm) | 72.46% | 70.22% | 97.16% | 80.20% |
This is an example that an IgG1 monoclonal antibody that was purified from cell culture medium using BioPro IEX SmartSep S30. In general, the purification of antibody starts with clarification. After clarification, the sample is subjected to initial purification (capture step) using affinity chromatography (rProtein A), followed by ion exchange chromatography. In the capture step, rProtein A derived from the affinity media contaminated the eluate, which was then separated and removed during ion exchange chromatography.
