Columns
BioPro HIC HT is a hydrophobic interaction chromatography column based on a non-porous hydrophilic polymer. It is suitable for analysis of biopharmaceuticals such as antibody-drug conjugates (ADC’s). Optimized surface modification and column packing condition realize high pressure tolerance and high separation ability. Therefore, high-throughput analysis with a faster flow rate can be achieved.
| SPECIFICATION | |
| Matrix | Hydrophilic non-porous polymer |
|---|---|
| Particle size | 2.3 µm |
| Bonded phase | Butyl group |
| Usable temp. range | 10-60°C |
| Usable pH range | 2-12 |
| Pressure limit | 20 MPa (33 X 4.6 mmI.D.) 40 MPa (100 X 4.6 mmI.D.) |
BioPro HIC HT consists of the fine non-porous polymer particle designed for separation of proteins, and has high pressure tolerance with excellent chromatographic resolution by optimized packing condition. This is capable of the rapid analysis without losing the resolution at a high pressure, which is too high to use with the other commercially available HIC column.
| Column | 100 X 4.6 mmI.D. |
|---|---|
| Eluent | A) 20 mM NaH2PO4-Na2HPO4 containing 1.0 M (NH4)2SO4 (pH 7.0) |
| Temperature | 25°C |
| Detection | UV at 280 nm |
| Injection | 10 µL |
| Sample | Brentuximab vedotin (2.5 mg/mL) |
Analysis throughput can be improved by using a 33 mm length column. This would contribute to rapid method development and improvement of cycle time. In addition, the column allows operation at a lower pressure.
| Column | BioPro HIC HT 2.3 µm |
|---|---|
| Eluent | A) 100 mM NaH2PO4-Na2HPO4 containing 2.3 M (NH4)2SO4 (pH 7.0) B) 100 mM NaH2PO4-Na2HPO4 (pH 7.0) |
| Temperature | 25°C |
| Detection | UV at 280 nm |
| Injection | 15 µL |
| Sample | 1. Ribonuclease A (0.75 mg/mL) 2. Lysozyme (0.25 mg/mL) 3. α-Chymotrypsinogen A (0.25 mg/mL) |
The DAR analysis by using BioPro HIC HT and the competitor columns under two mobile phase conditions varying 2-propanol content was carried out. The competitor A column having high hydrophobicity did not elute the peak #5 completely with the mobile phase containing 2-propanol lower than 15%. The competitor B column eluted all peaks with the 10% 2-propanol mobile phase condition, but it showed the poor separation between the peaks #4 and #5. On the other hand, BioPro HIC HT eluted all peaks completely with the good separation, and another peak was partially separated on the peak #3. Additionally, it is noteworthy that the same DAR values were observed at any content of 2-propanol.
| Column | 100 X 4.6 mmI.D. |
|---|---|
| Eluent | A) 20 mM NaH2PO4-Na2HPO4 containing 1.0 M (NH4)2SO4 (pH 7.0) |
| Flow rate | 0.5 mL/min |
| Temperature | 25°C |
| Detection | UV at 280 nm |
| Injection | 10 µL |
| Sample | Brentuximab vedotin (2.5 mg/mL) |
BioPro HIC HT exhibits an excellent batch-to-batch reproducibility. This is suitable for quality control analysis of biopharmaceuticals such as MAb’s.
| Column | BioPro HIC HT 2.3 µm, 100 X 4.6 mmI.D. |
|---|---|
| Eluent | A) 100 mM NaH2PO4-Na2HPO4 containing 2.0 M (NH4)2SO4 (pH 7.0) B) 100 mM NaH2PO4-Na2HPO4 (pH 7.0) 0%B (0-1 min), 0-100%B (1-11 min), 100%B (11-15 min) |
| Flow rate | 0.5 mL/min |
| Temperature | 25°C |
| Detection | UV at 280 nm |
| Injection | 15 µL |
| Sample | 1. Adalimumab (0.5 mg/mL) 2. Trastuzumab (0.5 mg/mL) 3. Bevacizumab (0.5 mg/mL) |
