Separation of Forced Aggregated mAb 1 Using Volatile and Salt-based Mobile Phase Compositions
| Columns: |
TSKgel UltraSW Aggregate, 3μm,
7.8 mm ID × 30 cm |
| Mobile phase: |
100 mmol/L PO4/100 mmol/L SO4,
pH 6.7
20% CH3CN/0.1% TFA/0.1% FA
100 mmol/L ammonium bicarbonate,
pH 7.0 |
| Gradient: |
isocratic
|
| Flow rate: |
1 mL/min
|
| Detection: |
UV @ 280 nm |
| Temperature: |
25 °C |
| Injection vol.: |
10 μL
|
| Sample: |
TBL mAb 1 (4.0 g/L) |
Separation of mAb Trimer and Dimer
| Column: |
TSKgel UltraSW Aggregate, 3 μm,
7.8 mm ID × 30 cm
|
| Mobile phase: |
0.2 mol/L phosphate buffer, pH 6.7 + 0.05% NaN3 |
| Flow Rate: |
0.8 mL/min |
| Detection: |
UV @ 280 nm |
| Temperature: |
25 °C
|
| Sample: |
monoclonal antibody-2
(mouse-human chimeric IgG, Erbitux), 10 μL |
|
|
Analysis of Heat Induced Forced Denatured, Large Hydrophobic Metalloprotein, Apoferritin
| Columns: |
TSKgel UltraSW Aggregate, 3 μm,
7.8 mm ID × 30 cm
|
| Mobile phase: |
50 mmol/L potassium phosphate (monobasic),
50 mmol/L sodium phosphate (dibasic),
100 mmol/L sodium sulfate, 0.05% NaN3, pH 6.7
|
| Flow rate: |
1.0 mL/min
|
| Detection: |
UV @ 280 nm
|
| Temperature: |
30 °C
|
| Injection vol.: |
10 μL
|
| Samples: |
ferritin - Sigma, 4.7 g/L, in saline (0.9% NaCl in water) solution, stored at 2-8°C
apoferritin - Sigma, 5.0 g/L, in 50% glycerol and 0.075 mol/L sodium chloride, stored at -20°C |
|
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