Analysis of mAb Aggregates
| Columns: |
A. TSKgel UP-SW Aggregate, 3 µm,
4.6 mm ID × 30 cm
B. TSKgel UP-SW3000, 2 µm,
4.6 mm ID × 30 cm |
| Mobile phase: |
40 mmol/L sodium phosphate buffer, pH 6.7
+ 400 mmol/L sodium perchlorate + 0.05% sodium azide |
| Flow rate: |
0.2 mL/min |
| Detection: |
UV @ 280 nm |
| Temperature: |
25 °C |
| Injection vol.: |
10 µL |
| Sample: |
mAb HMWs: dimer, aggregates; LMWs: fragments |
|
|
Analysis of Heat Forced mAb Aggregation
| Column: |
TSKgel UP-SW Aggregate, 3 µm,
4.6 mm ID × 30 cm |
| Mobile phase: |
40 mmol/L sodium phosphate buffer, pH 6.7,
+ 400 mmol/L sodium perchlorate + 0.05 % sodium azide |
| Flow rate: |
0.2 mL/min |
| Instruments: |
Agilent 1100 with Chemstation v. 2.18.18 |
| Detection: |
UV @ 280 nm |
| Temperature: |
25 °C |
| Injection vol.: |
10 µL |
| Sample: |
mAb |
* The sample was diluted by 10-fold with 20 mmol/L sodium phosphate buffer (pH 7.2) + 150 mmol/L sodium chloride and dispensed to small aliquots. Each aliquot was stored at following temperature respectively for 2 hours.
70 °C (black chromatogram)
73 °C (red chromatogram)
77 °C (green chromatogram)
80 °C (light blue chromatogram)
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